U.S. patent application number 16/625151 was filed with the patent office on 2021-11-11 for amino alcohol derivative, pharmaceutical composition and application thereof.
This patent application is currently assigned to Beijing Foreland Pharma Co., Ltd.. The applicant listed for this patent is Beijing Foreland Pharma Co., Ltd.. Invention is credited to Meijing Du, Zhenjian Du, Congmin Gao, Longlong Gong, Qi Ji, Lei Wang, Xingmin Zhang.
Application Number | 20210347745 16/625151 |
Document ID | / |
Family ID | 1000005765296 |
Filed Date | 2021-11-11 |
United States Patent
Application |
20210347745 |
Kind Code |
A1 |
Ji; Qi ; et al. |
November 11, 2021 |
AMINO ALCOHOL DERIVATIVE, PHARMACEUTICAL COMPOSITION AND
APPLICATION THEREOF
Abstract
The present invention belongs to the field of medicine, and
specifically discloses an amino alcohol derivative represented by
Formula I, a pharmaceutically acceptable salt, solvate, polymorph
or prodrug thereof. In addition, the present invention also
discloses a pharmaceutical composition comprising the above
substances, and a use of the substance in the preparation of a
medicament for the prevention and treatment of an immune
inflammatory disease, or a disease or condition associated with
immunological competence such as multiple sclerosis, ALS, CIDP,
systemic lupus erythematosus, rheumatoid arthritis, ulcerative
colitis, psoriasis, polymyositis, etc.
Inventors: |
Ji; Qi; (Yizhuang, Beijing,
CN) ; Zhang; Xingmin; (Yizhuang, Beijing, CN)
; Du; Zhenjian; (Yizhuang, Beijing, CN) ; Gong;
Longlong; (Yizhuang, Beijing, CN) ; Wang; Lei;
(Yizhuang, Beijing, CN) ; Gao; Congmin; (Yizhuang,
Beijing, CN) ; Du; Meijing; (Yizhuang, Beijing,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Beijing Foreland Pharma Co., Ltd. |
Yizhuang, Beijing |
|
CN |
|
|
Assignee: |
Beijing Foreland Pharma Co.,
Ltd.
Yizhuang, Beijing
CN
|
Family ID: |
1000005765296 |
Appl. No.: |
16/625151 |
Filed: |
June 21, 2018 |
PCT Filed: |
June 21, 2018 |
PCT NO: |
PCT/CN2018/092233 |
371 Date: |
December 20, 2019 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 45/06 20130101;
C07D 271/06 20130101; C07D 413/04 20130101 |
International
Class: |
C07D 271/06 20060101
C07D271/06; A61K 45/06 20060101 A61K045/06; C07D 413/04 20060101
C07D413/04 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 30, 2017 |
CN |
201710554216.6 |
Claims
1. An amino alcohol derivative represented by the following Formula
I, or a pharmaceutically acceptable salt, stereoisomer, isotopic
label, solvate, polymorph, or prodrug thereof: ##STR00084## wherein
R.sub.1 and R.sub.2 are the same or different, an is each
independently selected from H, --F, --Cl, --Br, --I, --OH, --SH,
--CN, --COOH, --NO.sub.2 and the following group of C.sub.1-40
alkyl, C.sub.1-40 alkoxy, C.sub.2-40 alkenyl, C.sub.2-40 alkynyl,
C.sub.3-20 cycloalkyl, C.sub.3-20 cycloalkoxy, 3- to 20-membered
heterocyclyl, 3- to 20-membered heterocycloxy, C.sub.6-20 aryl,
C.sub.6-20 aryloxy, 5- to 20-membered heteroaryl, 5- to 20-membered
heteroaryloxy, H[(CH.sub.2).sub.nO].sub.m--, --NR.sub.dR.sub.e,
--CONR.sub.dR.sub.e or --C(O)Y.sub.1R.sub.d, each of which is
unsubstituted or optionally substituted with one or more R.sub.a;
R.sub.3 is selected from the group consisting of C.sub.3-20
cycloalkyl, 3- to 20-membered heterocyclyl, C.sub.6-20 aryl and 5-
to 20-membered heteroaryl, each of which is unsubstituted or
optionally substituted with one or more R.sub.b; each R.sub.a is
the same as or different from any other one and is independently
selected from C.sub.1-40 alkyl, C.sub.1-40 alkoxy, C.sub.2-40
alkenyl, C.sub.2-40 alkynyl, C.sub.3-20 cycloalkyl, --F, --Cl,
--Br, --I, --OH, --NH, --SH, --CN, .dbd.O or --COOH; each R.sub.b
is the same as or different from any other one and is independently
selected from --F, --Cl, --Br, --I, --SH, --OH, --CN, --COOH and
the following group of C.sub.1-40 alkyl, C.sub.1-40 alkoxy,
C.sub.2-40 alkenyl, C.sub.2-40 alkynyl, C.sub.3-20 cycloalkyl, 3-
to 20-membered heterocyclyl, C.sub.6-20 aryl, 5- to 20-membered
heteroaryl, C.sub.3-20 cycloalkoxy, 3- to 20-membered
heterocycloxy, C.sub.6-20 aryloxy, 5- to 20-membered heteroaryloxy,
(C.sub.3-20)cycloalkyl(C.sub.1-40)alkyl, (3- to
20-membered)heterocyclyl(C.sub.1-40)alkyl,
(C.sub.6-20)aryl(C.sub.1-40)alkyl, (5- to
20-membered)heteroaryl(C.sub.1-40)alkyl,
H[(CH.sub.2).sub.nO].sub.n--, --NR.sub.cR.sub.d,
--C(O)NR.sub.cR.sub.d, --Y.sub.1C(O)R.sub.e or
--C(O)Y.sub.1R.sub.e, each of which is unsubstituted or optionally
substituted with one or more R.sub.a; or, when R.sub.3 is
substituted with two or more identical or different R.sub.b, two of
which losing their hydrogen atoms or other groups respectively, are
taken together with the carbon atoms to which they are attached to
form a ring system R.sub.s fused with R.sub.3, wherein R.sub.s is
selected from C.sub.3-20 cycloalkyl, 3- to 20-membered
heterocyclyl, C.sub.6-20 aryl, or 5- to 20-membered heteroaryl
fused with R.sub.3. R.sub.c, R.sub.d and R.sub.e are the same or
different, each of which is independently selected from H and the
following group of C.sub.1-40 alkyl, C.sub.2-40 alkenyl, C.sub.2-40
alkynyl, C.sub.3-20 cycloalkyl, 3- to 20-membered heterocyclyl,
C.sub.6-20 aryl, 5- to 20-membered heteroaryl or CONH.sub.2, each
of which is unsubstituted or optionally substituted with one or
more R.sub.a; Y.sub.1 is selected from a chemical bond, --O--,
--S--, and the group of --NH--, C.sub.1-40 alkyl, C.sub.1-40
alkoxy, C.sub.3-20 cycloalkyl, 3- to 20-membered heterocyclyl,
C.sub.6-20 aryl, 5- to 20-membered heteroaryl, or
(CH.sub.2CH.sub.2O).sub.j--, each of which is unsubstituted or
optionally substituted with one or more R.sub.a; m, n and j may be
the same or different, each of which is independently selected from
an integer equal to or more than 1, for example an integer in the
range of 1 to 20, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;
preferably, R.sub.1 and R.sub.2 may be the same or different, each
of which is independently selected from H, --F, --Cl, --Br, --I,
--OH, --SH, --CN, --COOH or C.sub.1-40 alkyl, for example, R.sub.1
or R.sub.2 is selected from H or C.sub.1-40 alkyl; R.sub.3 may be
selected from the group consisting of C.sub.3-8 cycloalkyl, 3- to
8-membered heterocyclyl, C.sub.6-10 aryl and 5- to 6-membered
heteroaryl, each of which is unsubstituted or optionally
substituted with one or more R.sub.b; each R.sub.b is the same as
or different from any other one and is independently selected from
--F, --Cl, --Br, --I, --SH, --OH, --CN, --COOH and the following
group of C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.3-8 cycloalkyl,
3- to 8-membered heterocyclyl, C.sub.6-10 aryl, 5- to 6-membered
heteroaryl, C.sub.3-8 cycloalkoxy, 3- to 8-membered heterocycloxy,
C.sub.6-10 aryloxy, 5- to 6-membered heteroaryloxy,
(C.sub.3-8)cycloalkyl(C.sub.1-6)alkyl, (3- to
8-membered)heterocyclyl(C.sub.1-6)alkyl,
(C.sub.6-10)aryl(C.sub.1-6)alkyl, (5- to
6-membered)heteroaryl(C.sub.1-6)alkyl,
H[(CH.sub.2).sub.nO].sub.n--, --NR.sub.cR.sub.d,
--C(O)NR.sub.cR.sub.d, --Y.sub.1C(O)R.sub.e or
--C(O)Y.sub.1R.sub.e, each of which is unsubstituted or optionally
substituted with one or more R.sub.a; or, when R.sub.3 is
substituted with two or more identical or different R.sub.b, two of
which losing their hydrogen atoms or other groups respectively, are
taken together with the carbon atoms to which they are attached to
form a ring system R.sub.s fused with R.sub.3, wherein R.sub.s is
selected from C.sub.3-8 cycloalkyl, 3- to 8-membered heterocyclyl,
C.sub.6-10 aryl, or 5- to 6-membered heteroaryl fused with
R.sub.3.
2. The amino alcohol derivative, or a pharmaceutically acceptable
salt, stereoisomer, isotopic label, solvate, polymorph, or prodrug
thereof according to claim 1, wherein R.sub.3 may be selected from
phenyl, pyridinyl, pyrazinyl, cyclohexyl, piperidinyl and
piperazinyl; for example, R.sub.3 may be selected from phenyl,
pyridin-1-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,
piperidin-1-yl, piperidin-2-yl, piperidin-3-yl and piperidin-4-yl;
preferably, each R.sub.b may be the same as or different from any
other one and is independently selected from --F, --Cl, --Br, --I,
--SH, --OH, --CN, --COOH and the following group of C.sub.1-6 alkyl
(such as methyl, ethyl, propyl, isopropyl, t-butyl), C.sub.1-6
alkoxy (methoxy, ethoxy, propoxy, i-propoxy, t-butoxy), C.sub.3-6
cycloalkyl, C.sub.3-6 cycloalkoxy, C.sub.1-6 alkylcarbonylamino,
C.sub.1-6 alkoxycarbonal, C.sub.1-6 alkylcarbonyloxy, (3- to
6-membered)heterocyclyl(C.sub.1-6)alkyl, --CONH.sub.2, and
--NHCOCH.sub.3, each of which is unsubstituted or optionally
substituted with one or more R.sub.a; for example, each R.sub.b may
be the same as or different from any other one and is independently
selected from --F, --OH, --CN, --CF.sub.3, --COOH, --CONH.sub.2,
methoxy, ethoxy, propoxy, i-propoxy, --NHCOCH.sub.3, cyclopentyl,
--C(O)OCH.sub.3, 1-azetidinylmethyl, 1-pyrrolidinylmethyl and
1-piperidinylmethyl; or, when R.sub.3 is substituted with two or
more identical or different R.sub.b, two of which losing their
hydrogen atoms or other groups respectively, are taken together
with the carbon atoms to which they are attached to form a ring
system R.sub.s fused with R.sub.3, wherein R.sub.s is selected from
dioxol ring system fused with R.sub.3; preferably, when R.sub.3 is
phenyl, which is preferably substituted with R.sub.b3 at least in
position 3 and R.sub.b3 is an electron withdrawing group;
preferably, when R.sub.3 is phenyl, which is preferably substituted
with R.sub.b4 at least in position 4 and R.sub.b4 is an electron
donating group; preferably, R.sub.b3 may be selected from --Cl,
--Br, --I, --SH, --OH, --CN, --COOH, --CONH.sub.2,
--CO--(C.sub.1-6)alkyl, --CO--(C.sub.3-6)cycloalkyl, and
--CF.sub.3; preferably, R.sub.b4 may be selected from C.sub.1-6
alkyl (such as methyl, ethyl, propyl, isopropyl, t-butyl),
C.sub.1-6 alkoxy (such as methoxy, ethoxy, propoxy, i-propoxy,
t-butoxy), C.sub.3-6 cycloalkyl, C.sub.3-6 cycloalkoxy, C.sub.1-6
alkylcarbonylamino.
3. The amino alcohol derivative, or a pharmaceutically acceptable
salt, stereoisomer, isotopic label, solvate, polymorph, or prodrug
thereof according to claim 1 or 2, wherein: the amino alcohol
derivative represented by Formula I has the structure of Formula
I': ##STR00085##
4. The amino alcohol derivative, or a pharmaceutically acceptable
salt, stereoisomer, isotopic label, solvate, polymorph, or prodrug
thereof according to any one of claims 1 to 3, wherein the compound
of Formula I is selected from the following compounds:
##STR00086##
5. A preparation method of the amino alcohol derivative, or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph, or prodrug thereof according to any one of
claims 1 to 4, comprising one or more steps of a to f: ##STR00087##
wherein R.sub.1, R.sub.2, and R.sub.3 are defined as any one of
claims 1 to 4; X is selected from halogen; PG.sub.1 is selected
from hydroxyl protecting groups; PG.sub.2 is selected from amino
protecting groups; or, PG.sub.1 may be linked with PG.sub.2 by a
bond so as to protect the hydroxyl group as well as the carbonyl
group.
6. The preparation method according to claim 5, wherein: in the
step a, D-tartaric acid may be used as a resolution agent to obtain
compound 7; in the step b, the amino group of the compound 7 may be
protected first in the presence of PhCHO and NaBH(OAc).sub.3, and
then compound 8 is obtained in the presence of
CH.sub.3OC(CH.sub.3).sub.2OCH.sub.3 (such as 2,2-dimethoxypropane)
and acid; in the step c, the compound 8 reacts in the presence of
n-BuLi and CO.sub.2 at the temperature of -78.degree. C. to obtain
compound 9; in the step d, the compound 9 may react with oxalyl
chloride first, then the resultant product reacts with compound 10
represented by the following formula in the presence of
triethylamine to obtain compound 11; ##STR00088## in the step e,
the compound 11 reacts in the presence of TsCl and triethylamine to
obtain compound 12; in the step f, the hydroxyl protecting group
PG.sub.1 and the amino protecting group PG.sub.2 are removed under
conditions for deprotection, for example, the hydroxyl protecting
group is removed in the presence of acid, and the amino protecting
group is removed under reduction condition; preferably, the
preparation method may comprise the following steps: ##STR00089##
##STR00090##
7. A pharmaceutical composition, comprising the amino alcohol
derivative, or a pharmaceutically acceptable salt, stereoisomer,
isotopic label, solvate, polymorph or prodrug thereof according to
any one of claims 1 to 4, and a pharmaceutically acceptable
carrier, wherein: the pharmaceutical composition may comprise, but
is not limited to, dosage forms for oral administration, parenteral
administration, topical administration and rectal administration;
preferably, the pharmaceutical composition is in the form of
tablet, capsule, pill, powder, sustained release preparation,
solution or suspension for oral administration; sterile solution,
suspension or emulsion for parenteral injection; ointment or cream
for external administration; or suppository for rectal
administration; preferably, the pharmaceutical composition is in a
unit dosage form suitable for single administration of a precise
dosage; preferably, the pharmaceutical composition further
comprises at least one therapeutic agent; preferably, the
pharmaceutical composition and the at least one therapeutic agent,
respectively, in separate dosage forms, are combined into a
combination product such as a kit of part.
8. Use of the amino alcohol derivative, or a pharmaceutically
acceptable salt, stereoisomer, isotopic label, solvate, polymorph
or prodrug thereof according to any one of claims 1 to 4, in the
preparation of a medicament for down-regulating the SIPl
expression.
9. Use of the amino alcohol derivative, or a pharmaceutically
acceptable salt, stereoisomer, isotopic label, solvate, polymorph
or prodrug thereof according to any one of claims 1 to 4, in the
preparation of a medicament for treating or preventing a disease or
condition associated with immune inflammation.
10. Use of the amino alcohol derivative, or a pharmaceutically
acceptable salt, stereoisomer, isotopic label, solvate, polymorph
or prodrug thereof according to any one of claims 1 to 4, in the
preparation of a medicament for treating or preventing a disease or
condition associated with immune activity, wherein: the disease or
condition associated with immune activity may be one or more of
multiple sclerosis, amyotrophic lateral sclerosis, CIDP, systemic
lupus erythematosus, rheumatoid arthritis, ulcerative colitis,
psoriasis, multiple myositis, type I diabetes, hyperthyroidism,
scleroderma and myasthenia gravis.
Description
TECHNICAL FIELD
[0001] The present application belongs to medical field,
particularly relates to an amino alcohol derivative, a
pharmaceutical composition and application thereof.
BACKGROUND
[0002] The immune system is a self-defensive structure mainly
consisting of lymphatic organs (thymus glands, lymph nodes, spleen,
tonsils), lymphatic tissues within other organs, lymphocytes
throughout the body, antigen presenting cell, and the like. The
immune system also includes other leucocytes in blood, and plasma
cells and mast cells in connective tissues. The key components of
the immune system are lymphocytes, which endue the immune system
with the capability of recognition and memory. The lymphocytes
travel throughout the body via blood and lymph, migrating from one
lymphatic organ or tissue to another lymphatic organ or tissue, and
connecting the lymphatic organs or tissues scattered throughout the
body to form a functional entirety. T cells and B cells are the
most important immunocytes in human bodies. The normal functioning
of each component of the immune system provides the guarantee for
the relative stability of the body immune functions, and any
deficiencies or hyperactions of the component would cause damage to
the body.
[0003] The components of immune system reach the whole body widely
and complicatedly, particularly with the continuous production,
circulation, and regeneration of the immune cells and immune
molecules. The immune system possesses a great recognizability,
which can precisely detect a foreign substance and distinguish it
from human's own healthy tissue in order to maintain body's
relative stability. Simultaneously, the immune system can accept,
transfer, enlarge, depot, and memorize the related immune
information, and provide positive or negative responses and
regulate the responsibility to the immune information. However, the
malfunctions of the immune system are disadvantageous to human
body: human's abnormal recognizability easily results in allergy
phenomenon, or causes iterative infections conversely; the abnormal
stabilizing ability may induce the immune system to give responses
to self-cells, which gives rise to autoimmune diseases.
[0004] Immunosuppressive agent is a type of new medicine category,
which developed from the foundation of the research on
neoplasm-chemotherapy, organ transplantation, immunopathology, and
clinical immunology, etc. It possesses immunosuppressive effects
which inhibits abnormal immune responses, and is generally used in
the therapy of organ transplant rejection and autoimmune diseases.
Common immunosuppressive agent comprises cyclophosphamide (CTX),
glucocorticoid, azathioprine, cyclosporine A (CsA), rapamycin,
mycophenolate mofetil and the like. Due to the restrictions of
selectivity and specificity, the above mentioned immunosuppressive
agents will inevitably damage immune defense capacity of the
patients when receiving the treatment, resulting in the descent
anti-infection ability of patients, the increasing risk of
malignant lesions, the injury of hematopoietic system, immune
system, liver, kidney and gastrointestinal function, neural and
endocrine function disorder, and inducing some allergic reactions,
etc. Therefore, the development and optimization of a new
immunosuppressive agent has become an important direction for new
drug development.
[0005] An agonist (such as FTY720) binds with the target molecule
of sphingosine-1-phosphate receptor 1 (S1P1, a kind of GPCR), which
leads to internalization of S1P1, down-regulation of the expression
of S1P1 on the surface of T lymphocytes and suppression of signal
transduction pathway of the target molecules. Therefore, the
inflammatory immune response mediated by activated T lymphocytes is
suppressed. The amino alcohol derivative has some structural
similarities with the endogenous hemolytic lipid of sphingosine.
Sphingosine is phosphorylated to form sphingosine-1-phosphate
induced by sphingosine enzyme. Activation of the receptor leads to
cells differentiation, growth and regulations of adhesion and
morphology of cells. In the normal immune responses, the
proliferation of T lymphocytes and B lymphocytes is taken place in
lymph nodes. They down-regulate the S1P1 expression when they are
in the lymph nodes. Once they are activated, the number of S1P1 on
cell surface will be up-regulated, which allowed T lymphocytes and
B lymphocytes to leave the lymph nodes. S1P1 on the surface of
lymphocytes may bind to drugs resulting in the down-regulation of
S1P1 expression, and thus losing the function of separating from
the lymph nodes. To the end, lymphocytes will adhere to the lymph
nodes. The amino alcohol derivative does not destroy the immune
function of the lymphocytes, but make the lymphocytes remained in
lymphatic system and inaccessible to blood circulatory system so
that the immune response is suppressed.
[0006] Sphingosine-1-phosphate receptor 1 agonist FTY720 has been
successfully developed by Novartis. However, FTY720 acts on not
only sphingosine-1-phosphate receptor 1 (S1P1), but
sphingosine-1-phosphate receptor 3 (S1P3), which can cause side
effects such as bradycardia. Therefore, the development of S1P1
agonist with better receptor selectivity is an important research
in this field.
SUMMARY OF THE INVENTION
[0007] In order to solve the technical problems in the prior art
mentioned above, the present invention provides an amino alcohol
derivative represented by the following Formula I, or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph, or prodrug thereof:
##STR00001##
[0008] wherein R.sub.1 and R.sub.2 are the same or different, and
is each independently selected from H, --F, --Cl, --Br, --I, --OH,
--SH, --CN, --COOH, --NO.sub.2 and the following group of
C.sub.1-40 alkyl, C.sub.1-40 alkoxy, C.sub.2-40 alkenyl, C.sub.2-40
alkynyl, C.sub.3-20 cycloalkyl, C.sub.3-20 cycloalkoxy, 3- to
20-membered heterocyclyl, 3- to 20-membered heterocycloxy,
C.sub.6-20 aryl, C.sub.6-20 aryloxy, 5- to 20-membered heteroaryl,
5- to 20-membered heteroaryloxy, H[(CH.sub.2).sub.nO].sub.m--,
--NR.sub.dR.sub.c, --CONR.sub.dR.sub.e or --C(O)Y.sub.1R.sub.d,
each of which is unsubstituted or optionally substituted with one
or more R.sub.a;
[0009] R.sub.3 is selected from the group consisting of C.sub.3-20
cycloalkyl, 3- to 20-membered heterocyclyl, C.sub.6-20 aryl and 5-
to 20-membered heteroaryl, each of which is unsubstituted or
optionally substituted with one or more R.sub.b;
[0010] each R.sub.a is the same as or different from any other one
and is independently selected from C.sub.1-40 alkyl, C.sub.1-40
alkoxy, C.sub.2-40 alkenyl, C.sub.2-40 alkynyl, C.sub.3-20
cycloalkyl, --F, --Cl, --Br, --I, --OH, --NH, --SH, --CN, .dbd.O or
--COOH;
[0011] each R.sub.b is the same as or different from any other one
and is independently selected from --F, --Cl, --Br, --I, --SH,
--OH, --CN, --COOH and the following group of C.sub.1-40 alkyl,
C.sub.1-40 alkoxy, C.sub.2-40 alkenyl, C.sub.2-40 alkynyl,
C.sub.3-20 cycloalkyl, 3- to 20-membered heterocyclyl, C.sub.6-20
aryl, 5- to 20-membered heteroaryl, C.sub.3-20 cycloalkoxy, 3- to
20-membered heterocycloxy, C.sub.6-20 aryloxy, 5- to 20-membered
heteroaryloxy, (C.sub.3-20)cycloalkyl(C.sub.1-40)alkyl, (3- to
20-membered)heterocyclyl(C.sub.1-40)alkyl,
(C.sub.6-20)aryl(C.sub.1-40)alkyl, (5- to
20-membered)heteroaryl(C.sub.1-40)alkyl,
H[(CH.sub.2).sub.nO].sub.n--, --NR.sub.cR.sub.d,
--C(O)NR.sub.cR.sub.d, --Y.sub.1C(O)R.sub.e and
--C(O)Y.sub.1R.sub.e, each of which is unsubstituted or optionally
substituted with one or more R.sub.a;
[0012] or, when R.sub.3 is substituted with two or more identical
or different R.sub.b, two of which losing their hydrogen atoms or
other groups respectively, are taken together with the carbon atoms
to which they are attached to form a ring system R.sub.s fused with
R.sub.3, wherein R.sub.s is selected from C.sub.3-20 cycloalkyl, 3-
to 20-membered heterocyclyl, C.sub.6-20 aryl, or 5- to 20-membered
heteroaryl fused with R.sub.3.
[0013] R.sub.c, R.sub.d and R.sub.e are the same or different, each
of which is independently selected from H and the following group
of C.sub.1-40 alkyl, C.sub.2-40 alkenyl, C.sub.2-40 alkynyl,
C.sub.3-20 cycloalkyl, 3- to 20-membered heterocyclyl, C.sub.6-20
aryl, 5- to 20-membered heteroaryl or CONH.sub.2, each of which is
unsubstituted or optionally substituted with one or more
R.sub.a;
[0014] Y.sub.1 is selected from a chemical bond, --O--, --S--, and
the group of --NH--, C.sub.1-40 alkyl, C.sub.1-40 alkoxy,
C.sub.3-20 cycloalkyl, 3- to 20-membered heterocyclyl, C.sub.6-20
aryl, 5- to 20-membered heteroaryl, or (CH.sub.2CH.sub.2O).sub.j--,
each of which is unsubstituted or optionally substituted with one
or more R.sub.a;
[0015] m, n and j may be the same or different, each of which is
independently selected from an integer equal to or more than 1, for
example an integer in the range of 1 to 20, such as 1, 2, 3, 4, 5,
6, 7, 8, 9 or 10.
[0016] According to one embodiment of the present invention,
wherein R.sub.1 and R.sub.2 may be the same or different, each of
which is independently selected from H, --F, --Cl, --Br, --I, --OH,
--SH, --CN, --COOH or C.sub.1-40 alkyl, for example, R.sub.1 or
R.sub.2 is selected from H or C.sub.1-40 alkyl;
[0017] R.sub.3 may be selected from the group consisting of
C.sub.3-8 cycloalkyl, 3- to 8-membered heterocyclyl, C.sub.6-10
aryl and 5- to 6-membered heteroaryl, each of which is
unsubstituted or optionally substituted with one or more
R.sub.b;
[0018] each R.sub.b is the same as or different from any other one
and is independently selected from --F, --Cl, --Br, --I, --SH,
--OH, --CN, --COOH and the following group of C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.3-8 cycloalkyl, 3- to 8-membered
heterocyclyl, C.sub.6-10 aryl, 5- to 6-membered heteroaryl,
C.sub.3-8 cycloalkoxy, 3- to 8-membered heterocycloxy, C.sub.6-10
aryloxy, 5- to 6-membered heteroaryloxy,
(C.sub.3-8)cycloalkyl(C.sub.1-6)alkyl, (3- to
8-membered)heterocyclyl(C.sub.1-6)alkyl,
(C.sub.6-10)aryl(C.sub.1-6)alkyl, (5- to
6-membered)heteroaryl(C.sub.1-6)alkyl,
H[(CH.sub.2).sub.nO].sub.n--, --NR.sub.cR.sub.d,
--C(O)NR.sub.cR.sub.d, --Y.sub.1C(O)R.sub.e or
--C(O)Y.sub.1R.sub.e, each of which is unsubstituted or optionally
substituted with one or more R.sub.a;
[0019] or, when R.sub.3 is substituted with two or more identical
or different R.sub.b, two of which losing their hydrogen atoms or
other groups respectively, are taken together with the carbon atoms
to which they are attached to form a ring system R.sub.s fused with
R.sub.3, wherein R.sub.s is selected from C.sub.3-8 cycloalkyl, 3-
to 8-membered heterocyclyl, C.sub.6-10 aryl, or 5- to 6-membered
heteroaryl fused with R.sub.3.
[0020] According to one embodiment of the present invention,
R.sub.3 may be selected from phenyl, pyridinyl, pyrazinyl,
cyclohexyl, piperidinyl and piperazinyl.
[0021] As an example, R.sub.3 may be selected from phenyl,
pyridin-1-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,
piperidin-1-yl, piperidin-2-yl, piperidin-3-yl and
piperidin-4-yl.
[0022] According to one embodiment of the present invention,
R.sub.3 could be substituted with each of R.sub.b at any suitable
position, such as at position 1, 2, 3, or 4 of R.sub.3.
[0023] According to one embodiment of the present invention, each
R.sub.b may be the same as or different from any other one and is
independently selected from --F, --Cl, --Br, --I, --SH, --OH, --CN,
--COOH and the following group of C.sub.1-6 alkyl (such as methyl,
ethyl, propyl, isopropyl, t-butyl), C.sub.1-6 alkoxy (methoxy,
ethoxy, propoxy, i-propoxy, t-butoxy), C.sub.3-6 cycloalkyl,
C.sub.3-6 cycloalkoxy, C.sub.1-6 alkylcarbonylamino, C.sub.1-6
alkoxycarbonal, C.sub.1-6 alkylcarbonyloxy, (3- to
6-membered)heterocyclyl(C.sub.1-6)alkyl, --CONH.sub.2, and
--NHCOCH.sub.3, each of which is unsubstituted or optionally
substituted with one or more R.sub.a.
[0024] As an example, each R.sub.b may be the same as or different
from any other one and is independently selected from --F, --OH,
--CN, --CF.sub.3, --COOH, --CONH.sub.2, methoxy, ethoxy, propoxy,
i-propoxy, --NHCOCH.sub.3, cyclopentyl, --C(O)OCH.sub.3,
1-azetidinylmethyl, 1-pyrrolidinylmethyl and
1-piperidinylmethyl;
[0025] or, when R.sub.3 is substituted with two or more identical
or different R.sub.b, two of which losing their hydrogen atoms or
other groups respectively, are taken together with the carbon atoms
to which they are attached to form a ring system R.sub.s fused with
R.sub.3, wherein R.sub.s is selected from dioxol ring system fused
with R.sub.3.
[0026] According to one embodiment of the present invention, when
R.sub.3 is phenyl, which is preferably substituted with R.sub.b3 at
least in position 3 and R.sub.b3 is an electron withdrawing
group.
[0027] According to a further embodiment of the present invention,
when R.sub.3 is phenyl, which is preferably substituted with
R.sub.N4 at least in position 4 and R.sub.N4 is an electron
donating group.
[0028] As an example, R.sub.b3 may be selected from --Cl, --Br,
--I, --SH, --OH, --CN, --COOH, --CONH.sub.2,
--CO--(C.sub.1-6)alkyl, --CO--(C.sub.3-6)cycloalkyl, and
--CF.sub.3.
[0029] As an example, R.sub.b4 may be selected from C.sub.1-6 alkyl
(such as methyl, ethyl, propyl, isopropyl, t-butyl), C.sub.1-6
alkoxy (such as methoxy, ethoxy, propoxy, i-propoxy, t-butoxy),
C.sub.3-6 cycloalkyl, C.sub.3-6 cycloalkoxy, C.sub.1-6
alkylcarbonylamino.
[0030] According to an amino alcohol derivative represented by
Formula I of the present invention, the amino alcohol derivative
may have the structure of Formula I':
##STR00002##
[0031] As an embodiment, the compound of Formula I in the present
invention may be selected from the following compounds:
TABLE-US-00001 Structure of Compounds ##STR00003## ##STR00004##
##STR00005## ##STR00006## ##STR00007## ##STR00008## ##STR00009##
##STR00010## ##STR00011## ##STR00012## ##STR00013## ##STR00014##
##STR00015## ##STR00016##
[0032] The present invention also provides a preparation method of
the amino alcohol derivative represented by Formula I comprising
one or more steps of a to f:
##STR00017##
[0033] wherein R.sub.1, R.sub.2, and R.sub.3 are defined as
previously;
[0034] X is selected from halogen;
[0035] PG.sub.1 is selected from hydroxyl protecting groups;
[0036] PG.sub.2 is selected from amino protecting groups;
[0037] or, PG.sub.1 may be linked with PG.sub.2 by a bond so as to
protect the hydroxyl group as well as the carbonyl group.
[0038] According to one embodiment of the preparation method in the
present invention, wherein:
[0039] in the step a, D-tartaric acid may be used as a resolution
agent to obtain compound 7;
[0040] in the step b, the amino group of the compound 7 may be
protected first in the presence of PhCHO and NaBH(OAc).sub.3, and
then compound 8 is obtained in the presence of
CH.sub.3OC(CH.sub.3).sub.2OCH.sub.3 and acid;
[0041] in the step c, the compound 8 reacts in the presence of
n-BuLi and CO.sub.2 at the temperature of -78.degree. C. to obtain
compound 9;
[0042] in the step d, the compound 9 may react with oxalyl chloride
first, then the resultant product reacts with compound 10
represented by the following formula in the presence of
triethylamine to obtain compound 11;
##STR00018##
[0043] in the step e, the compound 11 reacts in the presence of
TsCl and triethylamine to obtain compound 12;
[0044] in the step f, the hydroxyl protecting group PG.sub.1 and
the amino protecting group PG.sub.2 are removed under conditions
for deprotection, for example, the hydroxyl protecting group is
removed in the presence of acid, and the amino protecting group is
removed under reduction condition.
[0045] According to one embodiment of the preparation method in the
present invention, the method may comprise the following steps:
##STR00019## ##STR00020##
[0046] The present invention also provides a pharmaceutical
composition comprising an amino alcohol derivative represented by
Formula I of the present invention a pharmaceutically acceptable
salt, stereoisomer, isotopic label, solvate, polymorph or prodrug
thereof, and a pharmaceutically acceptable carrier.
[0047] According to the present invention, the pharmaceutical
composition may comprise, but is not limited to, dosage forms for
oral administration, parenteral administration, topical
administration and rectal administration.
[0048] In some embodiments, the pharmaceutical composition may be
in the form of tablet, capsule, pill, powder, sustained release
preparation, solution or suspension for oral administration;
sterile solution, suspension or emulsion for parenteral injection;
ointment or cream for topical administration; or suppository for
rectal administration.
[0049] In further embodiments, the pharmaceutical composition is in
a unit dosage form suitable for single administration of a precise
dosage.
[0050] In further embodiments, the amount of the compound is in a
range of about 0.001 mg/kg body weight/day to about 1000 mg/kg body
weight/day.
[0051] In further embodiments, the amount of the compound is in a
range of about 0.5 mg/kg body weight/day to about 50 mg/kg body
weight/day.
[0052] In some embodiments, the amount of the compound is about
0.001 g/day to about 7 g/day.
[0053] In further embodiments, the amount of the compound is about
0.002 g/day to about 6 g/day.
[0054] In further embodiments, the amount of the compound is about
0.005 g/day to about 5 g/day.
[0055] In further embodiments, the amount of the compound is about
0.01 g/day to about 5 g/day.
[0056] In further embodiments, the amount of the compound is about
0.02 g/day to about 5 g/day.
[0057] In further embodiments, the amount of the compound is about
0.05 g/day to about 2.5 g/day.
[0058] In further embodiments, the amount of the compound is about
0.1 g/day to about 1 g/day.
[0059] In further embodiments, dosage levels below the lower limit
of the aforesaid ranges may be adequate.
[0060] In further embodiments, dosage levels above the upper limit
of the aforesaid ranges may be required.
[0061] In some embodiments, the compound is administered in a
single dose once a day.
[0062] In further embodiments, the compound is administered in
multiple doses more than once a day.
[0063] In some embodiments, the compound is administered twice a
day.
[0064] In further embodiments, the compound is administered three
times a day.
[0065] In further embodiments, the compound is administered four
times a day.
[0066] In further embodiments, the compound is administered more
than four times a day.
[0067] In some embodiments, the individual to which the
pharmaceutical composition is administrated is a mammal.
[0068] In further embodiments, the mammal is human.
[0069] In further embodiments, the pharmaceutical composition
further comprises at least one therapeutic agent (i.e., formulated
into a single dosage form).
[0070] In some embodiments, the pharmaceutical composition and the
at least one therapeutic agent, respectively, in separate dosage
forms, are combined into a combination product such as a kit of
part.
[0071] The present invention also provides an amino alcohol
derivative represented by the Formula I described above or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph or prodrug thereof for use in the preparation of
a medicament for down-regulating S1P1 expression.
[0072] The present invention also provides an amino alcohol
derivative represented by the Formula I described above or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph or prodrug thereof for use in down-regulating
the S1P1 expression.
[0073] The present invention also provides a method for regulating
(e.g., down-regulating) the activity of S1P1, comprising contacting
S1P1 with an effective amount of the compound described above or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph or prodrug thereof.
[0074] Preferably, the method may be used in vivo, and may be also
used in vitro.
[0075] The present invention also provides an amino alcohol
derivative represented by the Formula I described above or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph or prodrug thereof for use in the preparation of
a medicament for treating or preventing a disease or condition
associated with immune inflammation.
[0076] The present application also provides an amino alcohol
derivative represented by the Formula I described above or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph or prodrug thereof for use in treating or
preventing a disease or condition associated with immune
inflammation.
[0077] The present application also provides an amino alcohol
derivative represented by the Formula I described above or a
pharmaceutically acceptable salt, stereoisomer, isotopic label,
solvate, polymorph or prodrug thereof for use in the preparation of
a medicament for treating or preventing a disease or condition
associated with immune activity.
[0078] The present application also provides a method for treating
a disease or condition associated with immune activity, comprising
administrating an effective amount of an amino alcohol derivative
represented by the Formula I described above or a pharmaceutically
acceptable salt, stereoisomer, isotopic label, solvate, polymorph
or prodrug thereof to an individual in need thereof.
[0079] According to the present invention, the individual may be a
mammal, such as human.
[0080] According to one embodiment of the present invention, the
disease or condition associated with immune activity may be one or
more of multiple sclerosis, amyotrophic lateral sclerosis, chronic
inflammatory demyelinating polyradiculoneuropathy (CIDP), systemic
lupus erythematosus, rheumatoid arthritis, ulcerative colitis,
psoriasis, multiple myositis, type I diabetes, hyperthyroidism,
scleroderma and myasthenia gravis.
Definition and Explanation of Terms
[0081] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as that commonly understood by
those skilled in the art to which the claimed subject belongs. All
patents, patent applications and published materials referred to
throughout the entire disclosure herein, unless noted otherwise,
are incorporated by reference in their entirety. In the event that
there is a plurality of definitions for terms herein, those in this
section prevail.
[0082] It should be understood that the foregoing general
description and the following detailed description are exemplary
and explanatory only and are not restrictive of any subject matter
claimed. In the present application, the use of the singular
includes the plural unless specifically stated otherwise. It should
also be noted that the use of "or" means "and/or" unless stated
otherwise. Besides, use of the term "comprising" as well as other
forms, such as "comprise", "comprises" and "comprised" is not
limiting.
[0083] Whenever a numerical range recited in the specification and
claims herein is defined as "an integer", it should be understood
that both endpoints of the range and each integer within the range
are recited. For example, "an integer of 0 to 10" should be
understood as reciting each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9
and 10. When the numerical range is defined as "a number", it
should be understood that both endpoints of the range, each integer
within the range, and each decimal within the range are recited.
For example, "a number from 0 to 10" should be understood as not
only each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also
at least the sum of each of the integers and 0.1, 0.2, 0.3, 0.4,
0.5, 0.6, 0.7, 0.8 and 0.9, respectively, are recited.
[0084] It should be understood that definition of standard chemical
terms may be found in reference works (including "Carey and
Sundberg `ADVANCED ORGANIC CHEMISTRY 4.sup.TH ED.` Vols. A (2000)
and B (2001), Plenum Press, New York"). Unless otherwise indicated,
conventional methods, such as mass spectrum, NMR, IR, and UV/Vis
spectroscopy and pharmacology methods, within the skill of the art
are employed. Unless specific definitions are provided, the
nomenclature employed in connection with the analytical chemistry,
synthetic organic chemistry, and medicinal and pharmaceutical
chemistry described herein are those known in the art. Standard
techniques can be used for chemical syntheses, chemical analyses,
pharmaceutical preparation, formulation, and delivery, and
treatment of patients. Reactions and purification techniques can be
performed e.g., using kits of manufacture's specifications or as
commonly accomplished in the art or as described herein. The
foregoing techniques and procedures can be generally performed by
conventional methods well known in the art and as described in
various general and more specific references that are cited and
discussed throughout the present specification. Throughout the
specification, groups and substituents thereof can be chosen by one
skilled in the art to provide stable moieties and compounds.
Wherein substituent groups are specified by their conventional
chemical formulas, written from left to right, they equally
encompass the chemically identical substituents that would result
from writing the structure from right to left. For example,
CH.sub.2O is equivalent to OCH.sub.2.
[0085] The term "optional" or "optionally" means that the
subsequently described event or circumstance may or may not occur,
and that the description includes instances wherein said event or
circumstance occurs and instances in which it does not.
[0086] The term "halogen" refers to F, Cl, Br and I. In other
words, F, Cl, Br and I could be described as halogen in the
specification.
[0087] It should be understood that the term "C.sub.1-40 alkyl"
preferably refers to a monovalent radical of a straight or branched
saturated hydrocarbon having 1 to 40 carbon atoms, and is
preferably C.sub.1-10 alkyl. It should be understood that
"C.sub.1-10 alkyl" preferably refers to a monovalent radical of a
straight or branched saturated hydrocarbon having 1, 2, 3, 4, 5, 6,
7, 8, 9 or 10 carbon atoms. The alkyl group is, for example,
methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl,
sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl,
1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl,
4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl,
2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl,
1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or
1,2-dimethylbutyl or isomer thereof. Specifically, the group has 1,
2, 3, 4, 5 or 6 carbon atoms ("C.sub.1-6 alkyl") such as methyl,
ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl.
More specifically, the group has 1, 2 or 3 carbon atoms ("C.sub.1-3
alkyl"), such as methyl, ethyl, n-propyl or isopropyl.
[0088] It should be understood that the term "C.sub.2-40 alkenyl"
preferably refers to a monovalent radical of a straight or branched
hydrocarbon containing one or more double bonds and 2 to 40 carbon
atoms, and is preferably C.sub.2-10 alkenyl. It should be
understood that the term "C.sub.2-10 alkenyl" preferably refers to
a monovalent radical of a straight or branched hydrocarbon
containing one or more double bonds and 2, 3, 4, 5, 6, 7, 8, 9 or
10 carbon atoms, especially 2 or 3 carbon atoms ("C.sub.2-3
alkenyl"). It should be understood when the alkenyl group comprises
more than one double bond, the double bonds could be conjugated or
isolated. The alkenyl group is, for example, vinyl, allyl,
(E)-2-methylethenyl, (Z)-2-methylethenyl, (E)-2-butenyl,
(Z)-2-butenyl, (E)-1-butenyl, (Z)-1-butenyl, 4-pentenyl,
(E)-3-pentenyl, (Z)-3-pentenyl, (E)-2-pentenyl, (Z)-2-pentenyl,
(E)-1-pentenyl, (Z)-1-pentenyl, 5-hexenyl, (E)-4-hexenyl,
(Z)-4-hexenyl, (E)-3-hexenyl, (Z)-3-hexenyl, (E)-2-hexenyl,
(Z)-2-hexenyl, (E)-1-hexenyl, (Z)-1-hexenyl, isopropenyl,
2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl,
(E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl,
3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl,
3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl,
(E)-1-methylbut-2-enyl, (Z)-1-methylbut-2-enyl,
(E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl,
(E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl,
(E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl,
1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylethenyl, or
1-isopropylethenyl.
[0089] It should be understood that the term "C.sub.2-40 alkynyl"
preferably refers to a monovalent radical of a straight or branched
hydrocarbon having one or more triple bonds and 2 to 40 carbon
atoms, and is preferably C.sub.2-10 alkynyl. It should be
understood that the term "C.sub.2-10 alkynyl" preferably refers to
a monovalent radical of a straight or branched hydrocarbon having
one or more triple bonds and 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon
atoms, especially 2 or 3 carbon atoms ("C.sub.2-3 alkynyl"). The
alkynyl group is, for example, ethynyl, 1-propynyl, 2-propynyl,
1-butynyl, 2-butynyl, 3-butynyl, 1-penynyl, 2-penynyl, 3-penynyl,
4-penynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl,
1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl,
1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl,
3-methylpen-4-ynyl, 2-methylpen-4-ynyl, 1-methylpen-4-ynyl,
2-methylpen-3-ynyl, 1-methylpen-3-ynyl, 4-methylpen-2-ynyl,
1-methylpen-2-ynyl, 4-methylpen-1-ynyl, 3-methylpen-1-ynyl,
2-ethylbut-3-ynyl, 1-ethylbut-3-ynyl, 1-ethylbut-2-ynyl,
1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl,
2,2-dimethylbut-3-ynyl, 1,1-dimethylbut-3-ynyl,
1,1-dimethylbut-2-ynyl, or 3,3-dimethylbut-1-ynyl. The alkynyl
group is especially ethynyl, 1-propynyl or 2-propynyl.
[0090] It should be understood that the term "C.sub.3-20
cycloalkyl" refers to a monovalent radical of a monocyclic or
bicyclic saturated hydrocarbon ring system having 3 to 20 carbon
atoms, and is preferably C.sub.3-10 cycloalkyl. It should be
understood that the term "C.sub.3-10 cycloalkyl" refers to a
monovalent radical of a monocyclic or bicyclic saturated
hydrocarbon ring system having 3, 4, 5, 6, 7, 8, 9 or 10 carbon
atoms. The C.sub.3-10 cycloalkyl may be a monocyclic hydrocarbon
group, for example, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl, or
may be a bicyclic hydrocarbon group such as decahydronaphthyl.
[0091] The term "3- to 20-membered heterocyclyl" refer to a
monovalent radical of a monocyclic or bicyclic saturated
hydrocarbon ring system having 1 to 5 ring heteroatoms, wherein
each heteroatom is independently selected from N, O, and S, and is
preferably 3- to 10-membered heterocyclyl. The term "3- to
10-membered heterocyclyl" refer to a monovalent radical of a
monocyclic or bicyclic saturated hydrocarbon ring system having 1
to 5, preferably 1 to 3 ring heteroatoms, wherein each heteroatom
is selected from N, O, and S. The heterocyclyl can be attached to
the rest moiety of a chemical structure through any ring carbon
atom or nitrogen atom (if present). Representative heterocyclyl
groups include, but are not limited to, 4-membered heterocyclyl,
such as azetidinyl, oxetanyl; 5-membered heterocyclyl, such as
terahydrofuryl, dioxolyl, pyrrolidinyl, imidazolidinyl,
pyrazolidinyl, pyrrolinyl; or 6-membered heterocyclyl, such as
tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl,
thiomorpholinyl, piperazinyl, or trithianyl; or 7-membered
heterocyclyl, such as diazepanyl. The heterocyclyl group may
optionally be a benzofused ring system. The heterocyclyl group may
be bicyclic, for example but not limited to 5,5-membered bicyclic
ring, such as hexahydrocyclopenta[c]pyrrol-2(1H)-yl, or
5,6-membered bicyclic ring, such as
hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl. The ring system containing
nitrogen atom(s) may be partially unsaturated, therefore it may
contain one or more double bonds, for example but is not limited to
2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl,
4,5-dihydrooxazolyl or 4H-[1,4]thiazinyl, or may be a benzofused
ring system, for example but is not limited to
dihydroisoquinolinyl, 1,3-benzoxazolyl, 1,3-benzodioxolyl.
According to the present invention, the heterocyclyl groups have no
aromaticity.
[0092] It should be understood that the term "C.sub.6-20 aryl"
refers to a monovalent radical of a mono-, bi-, or tri-cyclic
hydrocarbon ring system in which the whole or part of the structure
is aromatic, and is preferably "C.sub.6-14 aryl". It should be
understood that the term "C.sub.6-14 aryl" is preferably a
monovalent radical of a mono-, bi-, or tri-cyclic hydrocarbon ring
system in which the whole or part of the structure is aromatic,
having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms (C.sub.6-14
aryl). Specially, C.sub.6-14 aryl is a ring having 6 carbon atoms
(C.sub.6 aryl), such as phenyl, or biphenyl; or a ring having 9
carbon atoms (C.sub.9 aryl), such as indanyl or indenyl; or a ring
having 10 carbon atoms (C.sub.10 aryl), such as
tetrahydronaphthalenyl, dihydronaphthalenyl or naphthyl; or a ring
having 13 carbon atoms (C.sub.13 aryl), such as fluorenyl; or a
ring having 14 carbon atoms (C.sub.14 aryl), such as anthryl.
[0093] It should be understood that the term "5- to 20-membered
heteroaryl" comprises a monovalent radical of a mono-, bi-, or
tri-cyclic aromatic ring system having 5 to 20 ring atoms of which
1 to 5 ring atoms are independently selected from N, O and S, such
as "5- to 14-membered heteroaryl". It should be understood that the
term "5- to 14-membered heteroaryl" comprises a monovalent radical
of a mono-, bi-, or tri-cyclic aromatic ring system having 5, 6, 7,
8, 9, 10, 11, 12, 13, or 14 ring atoms, especially 5, 6, 9 or 10
carbon atoms, of which 1 to 5, preferably 1 to 3 ring atoms are
independently selected from N, O and S, and may be a benzofused
ring in any situation. The heteroaryl is especially selected from
thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl,
pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl,
thiadiazolyl, thia-4H-pyrazolyl and benzo derivatives thereof, for
example, benzofuranyl, benzothiophenyl, benzoxazolyl,
benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl,
isoindolyl; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl,
triazinyl and benzo derivatives thereof, for example, quinolyl,
quinazolinyl, isoquinolyl; or azocinyl, indolizinyl, purinyl and
benzo derivatives thereof, or cinnolinyl, phthalazinyl,
quinazolinyl, quinoxalinyl, naphthyridinyl, pteridyl, carbazolyl,
acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl and the
like.
[0094] Unless otherwise indicated, the heterocyclyl, heteroaryl, or
heteroarylene groups comprise all possible isomeric forms, such as
position isomers. Therefore, as non-limiting examples for
description, pyridyl or pyridylidene comprises 2-pyridyl,
2-pyridylidene, 3-pyridyl, 3-pyridylidene, 4-pyridyl and
4-pyridylidene; thienyl or thienylidene comprises 2-thienyl,
2-thienylidene, 3-thienyl and 3-thienylidene.
[0095] The definitions of the term "alkyl" above-mentioned, such as
"C.sub.1-40 alkyl", equally apply to the terms comprising
C.sub.1-40 alkyl, for example "C.sub.1-40 alkoxy", "C.sub.1-40
alkylsilyl", "C.sub.1-40 alkylsiloxy" and the like. Similarly, the
definitions above-mentioned of the terms "C.sub.2-40 alkenyl",
"C.sub.2-40 alkynyl", "C.sub.3-20 cycloalkyl", "C.sub.5-20
cycloalkenyl", "3- to 20-membered heterocyclyl", "C.sub.6-20 aryl"
and "5- to 20-membered heteroaryl" equally apply to the terms
comprising these terms, such as "C.sub.2-40 alkenoxy", "C.sub.2-40
alkynoxy", "C.sub.3-20 cycloalkoxy", "3- to 20-membered
heterocyclyl", "3- to 20-membered heterocycloxy", "C.sub.6-20
aryloxy", "C.sub.6-20 arylalkyl", "5- to 20-membered
heteroarylalkyl" and the like.
[0096] The term "protecting group" of the present invention refers
to a temporary substituent used for protecting a reactive group
against undesired chemical conversion. In any method for preparing
the compound of the present invention, it may be necessary and/or
desired to protect a sensitive or reactive group of any related
molecule, which may be performed relying on a known protecting
group, such as the protecting group described in a textbook or
reference book in the art. The protecting group may be removed in a
suitable step followed by using a known method in the art. It
should be known for those skilled in the art that other reagents
may be used in the deprotection step depending on the kind of the
protecting group, which comprise but not limited to Pd/C,
Pd(OH).sub.2, PdCl.sub.2, Pd(OAc).sub.2/Et.sub.3SiH, Raney nickel,
appropriate acid, appropriate base, fluoride and the like.
[0097] The relevant terms "subject", "patient" or "individual" as
used herein refer to an individual suffering from a disease,
disorder or condition, and encompasses mammals and non-mammals.
Examples of mammals include, but are not limited to, any member of
the mammalian class: humans, non-human primates such as chimpanzees
and other apes and monkeys; farm animals such as cattle, horses,
sheep, goats, swine; domestic animals such as rabbits, dogs, and
cats; laboratory animals including rodents, such as rats, mice and
guinea pigs, and the like. Examples of non-mammals include, but are
not limited to, birds, fishes, and the like. In one embodiment of
the method and composition provided herein, the mammal is a
human.
[0098] The terms "treat," "treating" or "treatment" as used herein
and other similar synonyms include alleviating, abating or
ameliorating a symptom of a disease or condition, preventing other
symptoms, ameliorating or preventing the underlying metabolic
causes of a symptom, inhibiting a disease or condition, e.g.,
arresting the development of the disease or condition, relieving
the disease or condition, causing improvement of the disease or
condition, relieving a symptom caused by the disease or condition,
or stopping a symptom of the disease or condition. In addition, the
term encompasses a purpose of prophylaxis. The term further
includes achieving a therapeutic benefit and/or a prophylactic
benefit. By therapeutic benefit it is meant to eradicate or
ameliorate the underlying disorder being treated. Furthermore, the
eradication or amelioration of one or more physiological symptoms
associated with the underlying disorder is also a therapeutic
benefit; for example, an improvement is observed in the patient,
notwithstanding that the patient may still be affected by the
underlying disorder. For the prophylactic benefit, the composition
may be administered to a patient at risk of developing a particular
disease, or to a patient reporting one or more physiological
symptoms of a disease, even though a diagnosis of this disease may
not have been made.
[0099] The terms "effective amount", "therapeutically effective
amount" or "pharmaceutically effective amount" as used herein refer
to a sufficient amount of at least one agent or compound being
administered which will relieve to some extent one or more symptoms
of the disease or condition being treated. The result can be
reduction and/or alleviation of the signs, symptoms or causes of a
disease, or any other desired alteration of a biological system.
For example, an "effective amount" for therapeutic uses is the
amount of the composition comprising a compound as disclosed herein
required to provide a clinically significant alleviation of a
disease. An effective amount suitable for any individual case may
be determined using techniques such as a dose escalation study.
[0100] The terms "administer", "administering", "administration",
or the like, as used herein, refer to a method that may be used to
deliver a compound or a composition to the desired site of
biological action. These methods include, but are not limited to,
oral route, transduodenal route, parenteral injection (including
intravenous, subcutaneous, intraperitoneal, intramuscular,
intra-arterial injection or infusion), topical and rectal
administration. A person skilled in the art is familiar with the
techniques for employing the compound and method described herein,
e.g., those discussed in Goodman and Gilman, The Pharmacological
Basis of Therapeutics, current ed.; Pergamon and Remington's,
Pharmaceutical Sciences (current edition), Mack Publishing Co.,
Easton, Pa. In preferred embodiments, the compound and composition
described herein are administered orally.
[0101] The term "acceptable" as used herein with respect to a
formulation, composition or ingredient means having no persistent
detrimental effect on the general health of the subject being
treated.
[0102] The term "pharmaceutically acceptable" as used herein,
refers to a material, such as a carrier or diluents, which does not
affect the biological activity or properties of the compounds
described herein, and is relatively nontoxic, i.e., the material
may be administered to an individual without causing undesirable
biological effects or interacting in a deleterious manner with any
of the components of the composition in which it is contained.
[0103] The term "pharmaceutical composition" as used herein, refers
to a biologically active compound, optionally mixed with at least
one pharmaceutically acceptable chemical component, such as, but
not limited to carriers, stabilizers, diluents, dispersing agents,
suspending agents, thickening agents, and/or excipients.
[0104] The term "carrier" as used herein, refers to relatively
nontoxic chemical compounds or agents that facilitate the
incorporation of a compound into cells or tissues.
[0105] The term "pharmaceutically acceptable salt" as used herein,
refers to salts that retain the biological effectiveness of the
free acids and bases of the specified compound and that are not
biologically or otherwise undesirable. The compound of the
invention also includes a pharmaceutically acceptable salt. The
pharmaceutically acceptable salt refers to those formed by
converting basic groups in the parent compound into a salt form.
The pharmaceutically acceptable salt includes, but not limited to,
inorganic or organic acid salts of the basic groups such as amine
(amino) groups. The pharmaceutically acceptable salt of the
invention may be synthesized from the parent compound, i.e., by
reaction of the basic groups in the parent compound with 1-4
equivalents of an acid in a solvent system. Suitable salts are
listed in Remington's Pharmaceutical Sciences, 17.sup.th ed., Mack
Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of
Pharmaceutical Science, 66, 2(1977), such as hydrochloride
salt.
[0106] Unless indicated specifically, the salt of the invention
refers to acidic salts formed by organic/inorganic acids, and basic
salts formed by organic/inorganic bases. In addition, when the
basic functional group in the compound of formula I is (but not
limited to) pyridine or imidazole, and the acidic function group is
(but not limited to) carboxylic acid, an amphoteric ion (an
inner-salt) will be formed. The inner-salts are also encompassed in
the salts of the invention.
[0107] The term "solvate" as used herein refers to a combination of
a compound of this invention with a solvent molecule formed by
solvation. In some embodiments, the solvate refers to a hydrate,
i.e., the solvent molecule is water molecule, and the combination
of a compound of this invention and water forms a hydrate. One or
more compounds of the invention may exist in the form of a solvate,
just like the solvates formed with the pharmaceutically acceptable
solvents such as water, ethanol, and the like. Therefore, the
invention includes both solvated and non-solvated forms. "Solvate"
refers to a physical aggregate formed with a compound of the
invention and one or more solvent molecules. This physical
aggregate includes different degrees of ions and covalent bonds,
for example, hydrogen bonds. It has been confirmed that this
solvate may be separated off, for example, when the lattice of a
crystal has one or more solvent molecules. "Solvate" includes both
parts of solvent phase and separable solvate. There are many
examples of the corresponding solvates, including ethanol solvate,
methanol solvate, and the like. "Hydrate" is a solvate in which the
solvate is water (H.sub.2O) molecule.
[0108] One or more compounds of the invention may be arbitrary
prepared into a solvate. The preparation of a solvate is well known
in the art. For example, the preparation of a solvate of the
antifungal drug, fluconazole, i.e., with ethyl acetate and water,
is described in M. Caira et al, J. Pharmaceutical Sci., 93(3),
601-611 (2004). Similar preparation methods of solvates and
hydrates are also described in E. C. van Tonder et al, AAPS
PharmSciTech., 5(1), article 12 (2004), and A. L. Bingham et al,
Chem. Commun., 603-604 (2001). A typical, non-limiting preparation
process is to dissolve the compound of the invention in a desired
amount of an ideal solvent (organic solvent or water or a mixture
thereof) at a temperature higher than normal temperature, cool,
stand and crystallize, and then separate off the crystals using a
standard method. The presence of the solvent (water) in the solvate
(hydrate) formed during the crystallization can be confirmed by an
I.R. spectroscopic analysis technology.
[0109] The terms "polymroph" or "polymrophism" as used herein refer
to a compound of this invention present in different crystal
lattice forms.
[0110] The term "an isotopic label" as used herein, refers to a
compound of the invention labelled by an isotope. For example, the
isotopes in the compound of the invention include various isotopes
of H, C, N, O, P, F, and S, such as .sup.2H, .sup.3H, .sup.13C,
.sup.14C, .sup.15N, .sup.18O, .sup.17O, .sup.31P, .sup.32P,
.sup.35S, .sup.18F and .sup.36S.
[0111] The term "pharmaceutically acceptable prodrug" as used
herein, refers to any pharmaceutically acceptable salt, ester, salt
of an ester or other derivative of a compound of this invention,
which, upon administration to a recipient, is capable of providing,
either directly or indirectly, a compound of this invention or a
pharmaceutically active metabolite or residue thereof. Particularly
preferred derivatives or prodrugs are those that increase the
bioavailability of the compounds of this invention when such
compounds are administered to a patient (e.g., by allowing orally
administered compound to be more readily absorbed into blood), or
those that enhance delivery of the parent compound to a biological
organ or action site (e.g., the brain or lymphatic system).
[0112] Various forms of prodrugs are well known in the art. See, T.
Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987)
Vol. 14 of the A.C.S. Symposium Series, Bioreversible Carriers in
Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical
Association and in Pergamon Press for discussion concerning
prodrugs. Design of Prodrugs, Bundgaard, A. Ed., Elseview, 1985 and
Method in Enzymology, Widder, K. et al., Ed.; Academic, 1985, vol.
42, p. 309-396; Bundgaard, H. "Design and Application of Prodrugs"
in A Textbook of Drug Design and Development, Krosgaard-Larsen and
H. Bundgaard, Ed., 1991, Chapter 5, p. 113-191; and Bundgaard, H.,
Advanced Drug Delivery Review, 1992, 8, 1-38, each of which is
incorporated herein by reference.
[0113] "Stereoisomer" as used herein, refers to isomers produced
from different arrangements of atoms in molecules in space. The
compound of formula I posses asymmetric or chiral centers, and thus
different stereoisomeric forms exist. All stereostructures of
molecular formula I, like a mixture, include racemic mixtures, as a
part of the present invention. Diastereomer isomers can be
separated into individual diastereomers, depending on their
different physicochemical properties, by using the well-known
means. For example, the resolution of individual enantiomers may be
achieved by reacting with suitable optically active substance (for
example, chiral alcohol or Mosher's acyl chloride) to convert into
diastereomers, and then separating them and converting (such as
hydrolyzing) into the corresponding individual isomers. Some
compounds of formula I may be atropisomers (such as substituted
aryl), which is also a part of the invention. Enantiomers may be
isolated by using a chiral chromatographic column. The compound of
formula I may exist different tautomeric forms, which are
encompassed in the scope of the invention, for example, compounds
in keto-enol and imine-enamine forms.
[0114] "The disease associated with immune activity" as used
herein, refers to a disease caused by immune problems mainly
including the following diseases: multiple sclerosis, amyotrophic
lateral sclerosis, CIDP, systemic lupus erythematosus, rheumatoid
arthritis, ulcerative colitis, psoriasis, multiple myositis, type I
diabetes, hyperthyroidism, scleroderma, myasthenia gravis and the
like.
Beneficial Effects of the Present Invention
[0115] The compound of the present invention can effectively target
sphingosine-1-phosphate receptor-1 (S1P1). The side effects, such
as bradycardia, are relieved when the compound of the present
invention target sphingosine-1-phosphate receptor-3 (S1P3). In
addition, the preparation method of the invention has a simple
process, mild reaction conditions and a high product yield.
DETAILED DESCRIPTION
[0116] Hereinafter, the compound of formula I, preparation method
and application thereof of the present invention will be described
in more detail through the examples. It is understood that the
following examples are merely exemplary descriptions and
explanations, and should not be construed as limiting the scope of
the present invention. Solutions obtained by a person skilled in
the art based on the contents above mentioned of the invention are
all covered in the scope of protection of the present
invention.
[0117] Unless otherwise indicated, the starting materials and
reagents in the following examples were all commercially available
products, or were prepared by the methods known in the art.
[0118] The conditions for LC-MS analysis in the synthesis process
are as follows:
[0119] Instrument: Agilent LCMS1260/MSD6120;
[0120] Chromatographic column: Agilent SB-C18, 2.1*50 mm, 1.8
.mu.m, SN: USWEY07289;
[0121] Mobile phase: A: H.sub.2O (0.1% FA) 90%, B: acetonitrile
10%, 0.400 ml/min, 45.00.degree. C.
[0122] Schedule
TABLE-US-00002 Time Function Parameters 2.24 Changing solvent
Solvent components A: 0.0% B: 100.0% components 3.00 Changing
solvent Solvent components A: 0.0% B: 100.0% components 3.01
Changing flow Flow rate: 0.5 ml/min rate 3.01 Changing solvent
Solvent components A: 90.0% B: 10.0% components 5.00 Changing
solvent Solvent components A: 90.0% B: 10.0% components 5.01
Changing flow Flow rate: 0.4 ml/min rate 5.01 Changing solvent
Solvent components A: 90.0% B: 10.0%. components
[0123] Instrument Parameters:
[0124] Ionization Mode: API-ES
[0125] Polarity: Positive
[0126] Collision-induced dissociations ascending order:
Disabled
[0127] Percentage of cycle time: 50.00%.
Preparation Example 1
##STR00021##
[0129] Synthesis of Intermediate 2
[0130] 100 g (465 mmol) of compound 1 was dissolved in 500 ml of
dichloromethane, cooled to 0.degree. C. in ice-salt bath, and added
dropwise with 120 g (930 mmol) of oxalyl chloride. After the
dropping was finished, the mixture obtained was heated at reflux
and reacted for 2 h. The reaction was monitored by TLC. After the
reaction was completed, the solvent was evaporated to dryness, and
dichloromethane was added and evaporated to dryness again to afford
112 g of intermediate, as a yellow liquid which was used directly
in the next step. 210 g (1410 mmol) of aluminum chloride was
suspended in 400 ml of dichloromethane, cooled to -10.degree. C. to
-5.degree. C., and then added dropwise with a solution of 112 g of
the above intermediate in 100 ml of dichloromethane. After the
dropping was finished, ethylene gas was introduced into the
reaction system for about 2 h keeping the temperature of
-10.degree. C. to -5.degree. C. After TLC showed that the reaction
was completed, the reaction solution was poured into ice-water
mixture and extracted with dichloromethane. The organic phases were
combined, washed twice with saturated sodium bicarbonate solution
and once with saturated sodium chloride solution, dried, and
evaporated to dryness to give a crude product, which was purified
with silica gel column (eluent: petroleum ether/ethyl acetate=5/1,
v/v) to give a product (85 g) as an orange-red liquid. Yield:
81.7%. LC-MS: 225, 227 [M+1].sup.+, t.sub.R=2.153 min.
[0131] Synthesis of Intermediate 3
[0132] Under the protection of nitrogen, 25.5 g (113 mmol) of
intermediate 2 was dissolved in 20 ml of dichloromethane, added
with 1.78 g (5.6 mmol) of zinc iodide, and added dropwise with 20.7
ml (167 mmol) of trimethylsilyl cyanide under cooling in a water
bath. The resulting mixture was stirred and reacted for 3 h at room
temperature. The reaction was monitored by TLC. After the reaction
was completed, a solution (80 ml, 20%) of ammonia in methanol was
added and stirred for 3 days at room temperature. The reaction was
stopped and the reaction solution was evaporated to dryness. A
solution (100 ml, 11%) of hydrogen chloride in methanol was added
to the residual. The mixture was stirred for 30 min, then added
with 400 ml of methyl tert-butyl ether, stirred for 30 minutes
again and filtered to give a product (27.9 g) as a light
yellowish-white solid. Yield: 85.5%. LC-MS: 251, 253 [M+1].sup.+,
t.sub.R=1.731 min.
[0133] Synthesis of Intermediate 4
[0134] 30 g (102 mmol) of intermediate 3 was suspended in 150 ml of
sulphuric acid (50%, v:v), heated to 150.degree. C. and reacted for
3 h. After HPLC showed that the reaction was completed, the mixture
obtained was cooled to room temperature, placed in a refrigerator
at 4.degree. C. overnight and filtered. The filter cake was washed
with concentrated hydrochloric acid and pumped to dryness in a
dryer to give a product (32 g), as a light brown solid. Yield:
100%. LC-MS: 270, 272 [M+1].sup.+, t.sub.R=1.382 min.
[0135] Synthesis of Intermediate 5
[0136] 12.1 g (102 mmol) of thionyl chloride was added to 200 ml of
methanol at 0.degree. C., stirred for 1 h, then added with 25 g (34
mmol) of intermediate 4, heated at reflux and reacted for 15 h. The
reaction was monitored by HPLC. After the reaction was completed,
the reaction solution was evaporated to dryness and adjusted pH to
8 with saturated sodium bicarbonate solution. The aqueous phase was
extracted with dichloromethane. The organic phases were combined,
dried and evaporated to dryness to give a product (9 g) as a
reddish brown oily substance. Yield: 91.1%. LC-MS: 284,286
[M+1].sup.+, t.sub.R=1.507 min.
Preparation of Intermediate 6:
(2-amino-7-bromo-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
[0137] 4.8 g (126 mmol) of lithium aluminium hydride was suspended
in 200 ml of tetrahydrofuran, cooled to -10.degree. C., added
dropwise with a solution of 18 g (63 mmol) of intermediate 5 in 100
ml of tetrahydrofuran and then reacted for 30 min at -10.degree. C.
The reaction was monitored by TLC. After the reaction was
completed, 4.8 ml of water, 14.4 ml of sodium hydroxide solution
(10%) and 24 ml of water were added sequentially to the reaction
system. The mixture was stirred for 20 min, then added with 70 g of
anhydrous sodium sulfate, stirred for 30 minutes and allowed to
stand overnight. The resulting mixture was filtered, dried and
evaporated to dryness to give a product (17 g) as a black oily
substance. Yield: 100%. LC-MS: 256, 258 [M+1].sup.+, t.sub.R=1.143
min.
Preparation of Intermediate 7-1:
(S)-(2-amino-7-bromo-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
[0138] The racemic mixture of intermediate 6 was separated by
recrystallization process with D-tartaric acid many times until
specific rotation of the product no longer increased to give chiral
intermediate 7-1. Specific Rotation [.alpha.].sub.D.sup.20=27 (C=1,
MeOH). LC-MS: 256,258 [M+1].sup.+, t.sub.R=1.143 min.
[0139] Synthesis of Intermediate 8-1
[0140] 17 g (66 mmol) of intermediate 7-1 and 7.75 g (73 mmol) of
benzaldehyde were dissolved in 200 ml of dichloromethane, added
with 6 g (99.6 mmol) of acetic acid, stirred for 1 hour, added with
21.1 g (99.6 mmol) of sodium triacetoxyborohyride in batches in an
ice-salt bath, and then reacted for 2 h at room temperature. The
reaction was monitored by TLC. After the reaction was completed,
the reaction system was adjusted to pH 8 with saturated sodium
bicarbonate solution. The aqueous phase was extracted with
dichloromethane. The organic phases were combined, dried and
evaporated to dryness to give a crude product, which was purified
with silica gel column (eluent: petroleum ether/ethyl acetate=10/1,
v/v) to give a product (15.6 g) as a black oily substance. Yield:
73%. LC-MS: 346, 348 [M+1].sup.+, t.sub.R=1.821 min.
[0141] Synthesis of Intermediate 8-2
[0142] 15.6 g (45 mmol) of intermediate 8-1 was dissolved in 120 ml
of 2,2-dimethoxypropane, added with 1 g (5.8 mmol) of
p-toluenesulfonic acid monohydrate and 10 g of molecular sieve,
heated to 135.degree. C. under confinement and reacted for 16 h.
The reaction was monitored by TLC. After the reaction was
completed, the resulting mixture was filtered, evaporated to
dryness, added with saturated sodium bicarbonate solution and
extracted with dichloromethane. The organic phases were combined,
dried and evaporated to dryness to give a crude product, which was
purified with silica gel column (eluent: petroleum ether/ethyl
acetate=30/1, v/v) to give a product (13 g) as a faint yellow
solid. Yield: 75.1%. LC-MS: 386, 388 [M+1].sup.+, t.sub.R=3.352
min.
[0143] Synthesis of Intermediate 9-1
[0144] 14 g (36 mmol) of intermediate 8-2 was dissolved in 100 ml
of tetrahydrofuran, cooled to -78.degree. C. under the protection
of nitrogen and added dropwise with 17 ml (43.5 mmol) of
n-butyllithium. The resulting mixture was stirred for 30 min at
-78.degree. C., then introduced with carbon dioxide gas for 30
minutes and naturally warmed to room temperature. The reaction was
monitored by TLC. After the reaction was completed, the reaction
solution was adjusted to pH 5-6 with acetic acid and evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=30/1, v/v) to give a
product (9.1 g) as a yellowish-white solid. Yield: 71.9%. LC-MS:
352 [M+1].sup.+, t.sub.R=2.415 min.
[0145] Synthesis of Intermediate 17
##STR00022##
[0146] 0.8 g (4.7 mmol) of material 16, 2.6 g (18.8 mmol) of
potassium carbonate and 1.8 ml (18.8 mmol) of 2-bromopropane were
suspended in 8 ml of DMF and reacted at 100.degree. C. for 1 h. The
reaction was monitored by TLC (PE/EA=3:1). After the reaction was
completed, 30 ml of saturated sodium bicarbonate solution and 30 ml
of ethyl acetate were added to the resulting mixture, which was
stirred, allowed to stand, and separated. The organic phase was
washed with 2.times.30 ml of water, dried and evaporated to dryness
to give a crude product, as a brown liquid (0.94 g). Yield:
93.7%.
[0147] Synthesis of Intermediate 19
##STR00023##
[0148] Under the protection of nitrogen, 0.94 g (4.4 mmol) of
intermediate 17 was suspended in 3.2 g (52.9 mmol, 85%) of
hydrazine hydrate, and reacted at 85.degree. C. for 500 min. The
reaction was monitored by TLC (PE/EA=1:1+Et.sub.3N). After the
reaction was completed, the resulting mixture was cooled to room
temperature, filtered, washed with water, and pumped to dryness to
give a white solid (0.86 g). Yield: 91.7%. LC-MS: 213 [M+1].sup.+,
t.sub.R=2.441 min.
[0149] Synthesis of Intermediate 21
##STR00024##
[0150] 0.9 g (2.56 mmol) of intermediate 9-1 was dissolved in 36 ml
of dichloromethane, added with 0.015 ml (cat.) of
N,N-dimethylformamide and cooled to 0.degree. C. 0.65 ml (7.68
mmol) of oxalyl chloride was added dropwise to the mixture, then
naturally warmed to room temperature and reacted for 1 h. After the
reaction was completed, the reaction solution was concentrated and
added with 16 ml of dichloromethane, as a stock solution. 0.82 g
(3.84 mmol) of compound 19 and 1.1 ml (7.68 mmol) of triethylamine
were dissolved in 16 ml of dichloromethane, cooled to 0.degree. C.,
added dropwise with the solution of acyl chloride in
dichloromethane obtained above, then naturally warmed to room
temperature and reacted overnight. The reaction was monitored by
TLC (PE/EA=3:1+AcOH). After the reaction was completed, 40 ml of
saturated sodium bicarbonate solution was added and the resulting
mixture was separated into layers. The aqueous phase was extracted
with dichloromethane. The organic phases were combined, dried and
rotary evaporated to dryness to give a crude product, which was
purified with silica gel column (eluent: petroleum ether/ethyl
acetate=1/1, v/v) to give a product (1.2 g) as a yellow solid.
Yield: 84.8%. LC-MS: 546 [M+1].sup.+, t.sub.R=4.125 min.
[0151] Synthesis of Intermediate 22
##STR00025##
[0152] 1.2 g (2.16 mmol) of intermediate 21 and 1 ml (6.49 mmol) of
triethylamine were dissolved in 24 ml of acetonitrile, cooled to
0.degree. C., added with 0.62 g (3.24 mmol) of 4-toluene sulfonyl
chloride, and stirred overnight at room temperature. The reaction
was monitored by TLC (PE/EA=1:1). After the reaction was completed,
water was added and the resulting mixture was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=10/1, v/v) to give a
product (0.7 g) as a white solid. Yield: 61.4%. LC-MS: 528
[M+1].sup.+, t.sub.R=5.226 min.
[0153] Synthesis of Intermediate 23
##STR00026##
[0154] 0.7 g (1.33 mmol) of intermediate 22 was dissolved in 3.7 ml
of hydrochloric acid (1 M) and 21 ml of methanol, and reacted for
120 min at 80.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated by rotary evaporation, added
with 20 ml of saturated sodium bicarbonate solution and 20 ml of
dichloromethane, and separated into layers. The aqueous phase was
extracted with dichloromethane. The organic phases were combined,
dried and rotary evaporated to dryness to give a crude product,
which was purified with silica gel column (eluent: petroleum
ether/ethyl acetate=1/2+1% Et.sub.3N, v/v) to give a product (0.6
g) as a white solid. Yield: 94.0%.
Example 1
(S)-(2-amino-7-(5-(3-fluoro-4-isopropoxyphenyl)-1,3,4-oxadiazol--
2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00027##
[0156] 0.6 g (1.24 mmol) of intermediate 23 was dissolved in 13 ml
of methanol, added with 0.01 ml of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of
0.122 g (20% m/m) of palladium 10% on carbon, purged with hydrogen,
and reacted for 240 min at 95.degree. C. The reaction was monitored
by TLC (DCM:MeOH=10:1). After the reaction was completed, the
resulting mixture was filtered, and the filter cake was washed with
plenty of methanol. The filtrate was concentrated by rotary
evaporation, added with 30 ml of saturated sodium bicarbonate
solution and 30 ml of dichloromethane, stirred, allowed to stand
and separated into layers. The aqueous phase was extracted with
dichloromethane. The organic phases were combined, dried, filtered
and rotary evaporated to give a crude product, which was purified
with silica gel column (eluent: DCM:MeOH=10/1+1% Et.sub.3N, V/V) to
give the target compound of Example 1 as a white solid (0.3 g).
Yield: 61.89%. LC-MS: 398 [M+1].sup.+, t.sub.R=3.090 min. A
corresponding hydrogenchloride salt was obtained by mixing the
solid with a solution of hydrogen chloride in methanol under
stirring.
Preparation Example 2
[0157] Synthesis of Intermediate 27
##STR00028##
[0158] 2.0 g (12 mmol) of intermediate 26 was dissolved in 20 ml of
methanol, cooled to 0.degree. C., added dropwise with 2.6 ml (36
mmol) of thionyl chloride, warmed to room temperature, stirred and
reacted overnight. The reaction was monitored by TLC. After the
reaction was completed, the reaction solution was adjusted to pH 8
with saturated sodium bicarbonate solution, and rotary evaporated
to dryness to remove methanol. The aqueous phase was extracted with
dichloromethane. The organic phases were combined, dried and rotary
evaporated to dryness to give a product (1.9 g) as a white solid.
Yield: 81.67%.
[0159] Synthesis of Intermediate 28
##STR00029##
[0160] Under the protection of nitrogen, 1.911 g (9.8 mmol) of
intermediate 27 was dissolved in 7 g (118.7 mmol, 85%) of hydrazine
hydrate, heated to 85.degree. C. and reacted for 7 h. The reaction
was monitored by TLC. After the reaction was completed, the
resulting mixture was cooled to room temperature, filtered and
pumped to dryness to give a white solid (1.728 g). Yield:
81.67%.
[0161] Synthesis of Intermediate 29
##STR00030##
[0162] 1.0 g (2.84 mmol) of intermediate 9-1 was dissolved in 40 ml
of dichloromethane, added with 5 drops of N,N-dimethylformamide and
cooled to 0.degree. C. 0.72 ml (8.52 mmol) of oxalyl chloride was
added dropwise to the mixture, then naturally warmed to room
temperature and reacted for 1 hour. After the reaction was
completed, the reaction solution was concentrated and added with 30
ml of dichloromethane, as a stock solution. 0.823 g (8.52 mmol) of
compound 28 and 1.19 ml (8.52 mmol) of triethylamine were dissolved
in 30 ml of dichloromethane and cooled to 0.degree. C. The solution
of acyl chloride in dichloromethane obtained above was added
dropwise to the mixture, then naturally warmed to room temperature
and reacted overnight. The reaction was monitored by TLC
(PE/EA=3:1+AcOH). After the reaction was completed, 500 ml of
saturated sodium bicarbonate solution was added and the resulting
mixture was separated into layers. The aqueous phase was extracted
with dichloromethane. The organic phases were combined, dried and
rotary evaporated to dryness to give a crude product, which was
purified with silica gel column (eluent: petroleum ether/ethyl
acetate=1/1, v/v) to give a product (1.3 g) as a yellow solid.
Yield: 89.08%. LC-MS: 514 [M+1].sup.+, t.sub.R=8.349 min.
[0163] Synthesis of Intermediate 30
##STR00031##
[0164] 1.3 g (2.53 mmol) of intermediate 29 and 1.06 ml (7.60 mmol)
of triethylamine were dissolved in 30 ml of acetonitrile, cooled to
0.degree. C., added with 0.73 g (3.80 mmol) of 4-toluene sulfonyl
chloride, and stirred overnight at room temperature. The reaction
was monitored by TLC (PE/EA=1:1). After the reaction was completed,
water was added and the resulting mixture was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=1/1, v/v) to give a
product (1.2 g) as a yellow solid. Yield: 95.65%.
[0165] Synthesis of Intermediate 31
##STR00032##
[0166] 1.2 g (2.42 mmol) of intermediate 30 was dissolved in 5.2 ml
of hydrochloric acid (1 M) and 30 ml of methanol, and reacted for
30 min at 80.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated by rotary evaporation, added
with 30 ml of saturated sodium bicarbonate solution and 30 ml of
dichloromethane, and separated into layers. The aqueous phase was
extracted with dichloromethane. The organic phases were combined,
dried and rotary evaporated to dryness to give a crude product,
which was purified with silica gel column (eluent: petroleum
ether/ethyl acetate=1/1+1% Et.sub.3N, v/v) to give a product (0.7
g) as a yellow solid. Yield: 63.64%.
Example 2
(S)-(2-amino-7-(5-(3-fluoro-4-isopropoxyphenyl)-1,3,4-oxadiazol--
2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00033##
[0168] 0.7 g (1.54 mmol) of intermediate 31 was dissolved in 30 ml
of methanol, added with 4 drops of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of
0.14 g (20% m/m) of palladium 10% on carbon, purged with hydrogen,
and reacted for 5 h at 95.degree. C. The reaction was monitored by
TLC (DCM:MeOH=10:1). After the reaction was completed, the
resulting mixture was filtered, and the filter cake was washed with
plenty of methanol. The filtrate was concentrated by rotary
evaporation, added with 30 ml of saturated sodium bicarbonate
solution and 30 ml of ethyl acetate, stirred, allowed to stand and
separated into layers. The aqueous phase was extracted with ethyl
acetate. The organic phases were combined, dried, filtered and
rotary evaporated to give a crude product, which was purified with
silica gel column (eluent: DCM:MeOH=10/1, V/V) to give the target
compound of Example 2 as a faint yellow solid (0.514 g). Yield:
91.35%. LC-MS: 366 [M+1].sup.+, t.sub.R=2.64 min. A corresponding
hydrogenchloride salt was obtained by mixing the solid with a
solution of hydrogen chloride in methanol under stirring. .sup.1H
NMR (400 MHz, DMSO) .delta. 8.31 (s, 3H), 7.89 (d, J=6.8 Hz, 2H),
7.77-7.68 (m, 1H), 7.63 (d, J=1.6 Hz, 1H), 7.37 (d, J=8.6 Hz, 1H),
7.15 (d, J=8.1 Hz, 1H), 6.18 (s, 2H), 3.17 (s, 2H), 3.08 (s, 2H),
3.02-2.81 (m, 3H), 2.11-1.90 (m, 2H).
Preparation Example 3
[0169] Synthesis of Intermediate 35
##STR00034##
[0170] Under the protection of nitrogen, 5.5 g (23.8 mmol) of
intermediate 34 and 2.3 g (26.1 mmol) of cuprous cyanide were
suspended in 30 ml of N-methylpyrrolidone, and reacted at
200.degree. C. for 5 h. The reaction was monitored by TLC. After
the reaction was completed, the resulting mixture was added with 60
ml of water and 60 ml of ethyl acetate, stirred for 30 min, and
filtered. The filter cake was washed with ethyl acetate and the
mother liquor was extracted with ethyl acetate. The organic phases
were combined, dried and rotary evaporated to dryness to give a
crude product, which was purified with silica gel column (eluent:
ethyl acetate/petroleum ether=1/1, v/v) to give a product (3.6 g)
as a yellow oily substance. Yield: 85.3%. LC-MS: 178 [M+1].sup.+,
t.sub.R=3.251 min.
[0171] Synthesis of Intermediate 36
##STR00035##
[0172] 3.6 g (20.3 mmol) of intermediate 35, 9.9 g (81.3 mmol) of
2-bromopropane and 11.2 g (81.3 mmol) of potassium carbonate were
suspended in 36 ml of N,N-dimethylformamide, and reacted at
90.degree. C. for 2 h. The reaction was monitored by TLC. After the
reaction was completed, the resulting mixture was added with 30 ml
of water and 30 ml of ethyl acetate, and separated into layers. The
organic phase was washed with water for 3 times, dried and rotary
evaporated to dryness to give a crude product (2.6 g) as a yellow
oily substance. Yield: 58.1%.
[0173] Synthesis of Intermediate 37
##STR00036##
[0174] Under the protection of nitrogen, 2.6 g (11.8 mmol) of
intermediate 36 was dissolved in 5.9 g (110 mmol, 85%) of hydrazine
hydrate, and reacted at 60.degree. C. for 1 h. The reaction was
monitored by TLC. After the reaction was completed, the resulting
mixture was cooled to room temperature, filtered, washed with
water, and pumped to dryness to give a white solid (2 g). Yield:
77.1%.
[0175] Synthesis of Intermediate 38
##STR00037##
[0176] 2.6 g (7.6 mmol) of intermediate 9-1 was dissolved in 120 ml
of dichloromethane, added with 0.01 g (cat.) of
N,N-dimethylformamide and cooled to 0.degree. C. 2.9 g (22.8 mmol)
of oxalyl chloride was added dropwise to the mixture, then
naturally warmed to room temperature and reacted for 3 h. The
reaction was monitored by TLC. After the reaction was completed,
the reaction solution was concentrated and added with 50 ml of
dichloromethane, as a stock solution. 2 g (9.1 mmol) of compound 37
and 7.9 g (22.8 mmol) of triethylamine were dissolved in 50 ml of
dichloromethane and cooled to 0.degree. C. The solution of acyl
chloride in dichloromethane obtained above was added dropwise to
the mixture, then naturally warmed to room temperature and reacted
overnight. The reaction was monitored by TLC. After the reaction
was completed, 100 ml of saturated sodium bicarbonate solution was
added and the resulting mixture was separated into layers. The
aqueous phase was extracted with dichloromethane. The organic
phases were combined, dried and rotary evaporated to dryness to
give a crude product, which was purified with silica gel column
(eluent: petroleum ether/ethyl acetate=1/1, v/v) to give a product
(4.1 g) as a yellow solid. Yield: 97.3%.
[0177] Synthesis of Intermediate 39
##STR00038##
[0178] 4.1 g (7.4 mmol) of intermediate 38 and 2.2 g (22.2 mmol) of
triethylamine were dissolved in 120 ml of acetonitrile, added with
2.1 g (11.1 mmol) of 4-toluene sulfonyl chloride, and stirred
overnight at room temperature. The reaction was monitored by TLC.
After the reaction was completed, water was added and the resulting
mixture was separated into layers. The aqueous phase was extracted
with dichloromethane. The organic phases were combined, dried and
evaporated to dryness to give a crude product (4.3 g) as a tan
solid. Yield: 100%.
[0179] Synthesis of Intermediate 40
##STR00039##
[0180] 4.3 g (8 mmol) of intermediate 39 was dissolved in 20.1 ml
of hydrochloric acid (1 M) and 120 ml of methanol, and reacted for
2 h at 80.degree. C. The reaction was monitored by TLC. After the
reaction was completed, 50 ml of saturated sodium bicarbonate
solution was added and the resulting mixture was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=1/1, v/v) to give a
product (1.5 g) as a white solid. Yield: 37.5%.
Example 3
(S)-5-(5-(7-amino-7-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-
-2-yl)-1,3,4-oxadiazol-2-yl)-2-isopropoxybenzonitrile
##STR00040##
[0182] 1.5 g (3.0 mmol) of intermediate 40 was dissolved in 45 ml
of methanol, added with 0.5 ml of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of 0.3
g (20% m/m) of palladium 10% on carbon, purged with hydrogen, and
reacted for 8 h at 80.degree. C. The reaction was monitored by TLC.
After the reaction was completed, the resulting mixture was
filtered, and the filter cake was washed with hot methanol. The
filtrate was rotary evaporated to dryness to give a crude product
(1.5 g), which was recrystallized in 15 ml of methanol, filtered
and washed to give the target compound of Example 3 as a white
solid (0.65 g). Yield: 54.1%. LC-MS: 405 [M+1].sup.+, t.sub.R=3.002
min. A corresponding hydrogenchloride salt was obtained by mixing
the solid with a solution of hydrogen chloride in methanol under
stirring. .sup.1H NMR (400 MHz, DMSO) .delta. 8.51 (d, J=2.2 Hz,
1H), 8.37 (dd, J=9.0, 2.2 Hz, 1H), 8.12 (s, 3H), 7.96 (d, J=6.6 Hz,
2H), 7.54 (d, J=9.2 Hz, 1H), 7.40 (d, J=8.6 Hz, 1H), 5.62 (t, J=5.1
Hz, 1H), 5.01-4.87 (m, 1H), 3.47 (d, J=4.9 Hz, 2H), 3.22-2.83 (m,
5H), 2.06-1.89 (m, 2H), 1.38 (d, J=6.0 Hz, 6H).
Example 4
(S)-5-(5-(7-amino-7-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-
-2-yl)-1,3,4-oxadiazol-2-yl)-2-isopropoxybenzamide
##STR00041##
[0184] 0.2 g (0.49 mmol) of the target compound of Example 3 was
dissolved in 5 ml of dimethyl sulfoxide, added with 0.25 g (1.8
mmol) of potassium carbonate, cooled to 0.degree. C., added
dropwise with 2 ml of hydrogen peroxide solution (30%), and stirred
for 2 h at room temperature. The reaction was monitored by TLC.
After the reaction was completed, the resulting solution was added
with 30 ml of water, stirred for 10 min, filtered, washed with
water, and pumped to dryness to give a crude product. The crude
product was recrystallized in 20 ml of methanol, filtered and
washed to give the target compound of Example 4 as a white solid
(110 mg). Yield: 53.4%. A corresponding hydrogenchloride salt was
obtained by mixing the solid with a solution of hydrogen chloride
in methanol under stirring. H NMR (400 MHz, DMSO) .delta. 8.50 (d,
J=2.3 Hz, 1H), 8.26 (s, 3H), 8.19 (dd, J=8.8, 2.3 Hz, 1H), 7.90 (s,
2H), 7.79 (s, 1H), 7.65 (s, 1H), 7.40 (t, J=9.5 Hz, 2H), 6.11-5.14
(br s, 1H), 5.01-4.85 (m, 1H), 3.48 (s, 3H), 3.19-2.76 (m, 4H),
2.00 (t, J=6.4 Hz, 2H), 1.40 (d, J=6.0 Hz, 6H).
Example 5
(S)-5-(5-(7-amino-7-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-
-2-yl)-1,3,4-oxadiazol-2-yl)-2-hydroxybenzonitrile
##STR00042##
[0186] 100 mg (0.24 mmol) of the target compound of Example 3 was
dissolved in sulphuric acid (50%), and reacted for 1 h at
90.degree. C. The reaction was monitored by TLC. After the reaction
was completed, the resulting mixture was added with saturated
sodium bicarbonate solution to adjust pH to 2-3, filtered, washed
with water, and pumped to dryness to give the target compound of
Example 5 as a white solid (35 mg). Yield: 34.3%. LC-MS: 363
[M+1].sup.+, t.sub.R=1.142 min. A corresponding hydrogenchloride
salt was obtained by mixing the solid with a solution of hydrogen
chloride in methanol under stirring.
Preparation Example 4
[0187] Synthesis of Intermediate 46
##STR00043##
[0188] 2.0 g (9.08 mmol) of material 45, 0.98 g (18.17 mmol) of
sodium methoxide and a small amount of TBAB were suspended in 40 ml
of THF, stirred for 30 min at room temperature, followed by the
addition of 1.7 ml of methyl iodide, stirred and reacted at room
temperature. The reaction was monitored by TLC (PE/EA=3:1). The
reaction solution was added with methyl iodide until the material
was reacted completely, then concentrated by rotary evaporation,
added with 40 ml of saturated sodium bicarbonate solution and 40 ml
of dichloromethane, stirred, allowed to stand and separated. The
aqueous phase was extracted with 2.times.40 ml of dichloromethane.
The organic phases were combined, dried and rotary evaporated to
dryness to give a crude product (2.1 g) as a yellow solid. Yield:
100%. LC-MS: 235 [M+1].sup.+, t.sub.R=4.457 min.
[0189] Synthesis of Intermediate 47
##STR00044##
[0190] Under the protection of nitrogen, 2.12 g (9.08 mmol) of
intermediate 46 was suspended in 6.4 ml (108.96 mmol, 85%) of
hydrazine hydrate, and reacted at 85.degree. C. for 100 min. The
reaction was monitored by TLC (PE/EA=1:1+Et.sub.3N). After the
reaction was completed, the resulting mixture was cooled to room
temperature, filtered, washed with water, and pumped to dryness to
give a crude product (1.96 g), as a white solid. Yield: 92.4%.
LC-MS: 235 [M+1].sup.+, t.sub.R=3.005 min.
[0191] Synthesis of Intermediate 48
##STR00045##
[0192] 2.9 g (8.25 mmol) of intermediate 9-1 was dissolved in 60 ml
of dichloromethane, added with 0.01 g (cat.) of
N,N-dimethylformamide and cooled to 0.degree. C. 2.1 ml (24.75
mmol) of oxalyl chloride was added dropwise to the mixture, then
naturally warmed to room temperature and reacted for 1 h. After the
reaction was completed, the reaction solution was concentrated and
added with 30 ml of dichloromethane, as a stock solution. 2.32 g
(9.9 mmol) of compound 47 and 3.5 ml (24.75 mmol) of triethylamine
were dissolved in 30 ml of dichloromethane and cooled to 0.degree.
C. The solution of acyl chloride in dichloromethane obtained above
was added dropwise to the mixture, then naturally warmed to room
temperature and reacted for 3 h. The reaction was monitored by TLC
(PE/EA=1:1+AcOH). After the reaction was completed, 70 ml of
saturated sodium bicarbonate solution was added and the resulting
mixture was separated into layers. The aqueous phase was extracted
with dichloromethane. The organic phases were combined, dried and
rotary evaporated to dryness to give a crude product, which was
purified with silica gel column (eluent: petroleum ether/ethyl
acetate=1/1, v/v) to give a product (3.1 g) as a brown solid.
Yield: 66.2%. LC-MS: 568 [M+1], t.sub.R=4.678 min.
[0193] Synthesis of Intermediate 49
##STR00046##
[0194] 3.1 g (5.46 mmol) of intermediate 48 and 2.3 ml (16.38 mmol)
of triethylamine were dissolved in 124 ml of acetonitrile, cooled
to 0.degree. C., added with 1.56 g (8.19 mmol) of 4-toluene
sulfonyl chloride, and stirred overnight at room temperature. The
reaction was monitored by TLC (PE/EA=1:1). After the reaction was
completed, water was added and the resulting mixture was separated
into layers. The aqueous phase was extracted with dichloromethane.
The organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=3/1, v/v) to give a
product (1.9 g) as a faint yellow solid. Yield: 63.3%. LC-MS: 550
[M+1].sup.+, t.sub.R=6.464 min.
[0195] Synthesis of Intermediate 50
##STR00047##
[0196] 1.0 g (1.82 mmol) of intermediate 49 was dissolved in 4.55
ml of hydrochloric acid (1 M) and 30 ml of methanol, and reacted
for 100 min at 80.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated by rotary evaporation, added
with 50 ml of saturated sodium bicarbonate solution and 50 ml of
dichloromethane, and separated into layers. The aqueous phase was
extracted with dichloromethane. The organic phases were combined,
dried and rotary evaporated to dryness to give a crude product,
which was purified with silica gel column (eluent: petroleum
ether/ethyl acetate=1/2+1% Et.sub.3N, v/v) to give a product (0.8
g) as a white solid. Yield: 86.32%.
Example 6
(S)-(2-amino-7-(5-(4-methoxy-3-(trifluoromethyl)phenyl)-1,3,4-ox-
adiazol-2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00048##
[0198] 0.8 g (1.57 mmol) of intermediate 50 was dissolved in 16 ml
of methanol, added with 0.1 ml of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of
0.16 g (20% m/m) of palladium 10% on carbon, purged with hydrogen,
and reacted for 5 h at 95.degree. C. The reaction was monitored by
TLC (DCM:MeOH=10:1). After the reaction was completed, the
resulting mixture was filtered, and the filter cake was washed with
hot methanol. The filtrate was rotary evaporated to dryness to give
a crude product, which was recrystallized in 24 ml of methanol,
filtered and washed to give the target compound of Example 6 as a
white solid (0.36 g). Yield: 54.67%. LC-MS: 420 [M+1].sup.+,
t.sub.R=3.002 min. A corresponding hydrogenchloride salt was
obtained by mixing the solid with a solution of hydrogen chloride
in methanol under stirring.
Preparation Example 5
[0199] Synthesis of Intermediate 54
##STR00049##
[0200] Under the protection of nitrogen, 2.0 g (11.62 mmol) of
material 53 was dissolved in 20 ml of methanol, added with 8.2 ml
(139.44 mmol, 85%) of hydrazine hydrate, and reacted at room
temperature for 1.5 h. The reaction was monitored by TLC
(PE/EA=3:1). After the reaction was completed, the resulting
mixture was added with 50 ml of water and stirred to precipitate
out a solid, filtered, washed with water, and pumped to dryness.
The aqueous phase was extracted with EA. The organic phase was
dried, concentrated by rotary evaporation, and pumped to dryness to
give a product (2 g), as a white solid. Yield: 100%. LC-MS: 173
[M+1].sup.+, t.sub.R=2.528 min.
[0201] Synthesis of Intermediate 56
##STR00050##
[0202] 1 g (2.84 mmol) of intermediate 9-1 was dissolved in 40 ml
of dichloromethane, added with 0.01 g (cat.) of
N,N-dimethylformamide and cooled to 0.degree. C. 0.72 ml (8.52
mmol) of oxalyl chloride was added dropwise to the mixture, then
naturally warmed to room temperature and reacted for 1 hour. After
the reaction was completed, the reaction solution was concentrated
and added with 20 ml of dichloromethane, as a stock solution. 0.54
g (3.13 mmol) of compound 54 and 1.2 ml (8.52 mmol) of
triethylamine were dissolved in 20 ml of dichloromethane and cooled
to 0.degree. C. The solution of acyl chloride in dichloromethane
obtained above was added dropwise to the mixture, then naturally
warmed to room temperature and reacted overnight. The reaction was
monitored by TLC (PE/EA=3:1+AcOH). After the reaction was
completed, 40 ml of saturated sodium bicarbonate solution was added
and the resulting mixture was separated into layers. The aqueous
phase was extracted with dichloromethane. The organic phases were
combined, dried and rotary evaporated to dryness to give a crude
product, which was purified with silica gel column (eluent:
petroleum ether/ethyl acetate=1/1, v/v) to give a product (0.7 g)
as a yellow solid. Yield: 48.8%. LC-MS: 506 [M+1].sup.+,
t.sub.R=4.587 min.
[0203] Synthesis of Intermediate 57
##STR00051##
[0204] 0.7 g (1.38 mmol) of intermediate 56 and 0.6 ml (4.14 mmol)
of triethylamine were dissolved in 38 ml of acetonitrile, cooled to
0.degree. C., added with 0.32 g (1.66 mmol) of 4-toluene sulfonyl
chloride, and stirred for 5 h at room temperature. The reaction was
monitored by TLC (PE/EA=1:1). After the reaction was completed,
water was added and the resulting mixture was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=10/1, v/v) to give a
product (0.43 g) as a white solid. Yield: 63.9%. LC-MS:
488[M+1].sup.+, t.sub.R=6.464 min.
[0205] Synthesis of Intermediate 58
##STR00052##
[0206] 0.4 g (0.82 mmol) of intermediate 57 was dissolved in 2.1 ml
of hydrochloric acid (1 M) and 16 ml of methanol, and reacted for 1
h at 80.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, 40 ml of
saturated sodium bicarbonate solution was added and the resulting
mixture was separated into layers. The aqueous phase was extracted
with dichloromethane. The organic phases were combined, dried and
rotary evaporated to dryness to give a crude product, which was
purified with silica gel column (eluent: petroleum ether/ethyl
acetate=1/1+1% Et.sub.3N, v/v) to give a product (0.242 g) as a
white solid. Yield: 66.0%.
Example 7
(S)-(2-amino-7-(5-(3,4-difluorophenyl)-1,3,4-oxadiazol-2-yl)-1,2-
,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00053##
[0208] 0.24 g (0.54 mmol) of intermediate 58 was dissolved in 8 ml
of methanol, added with 0.01 ml of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of
0.048 g of palladium 10% on carbon, purged with hydrogen, and
reacted for 5 h at 95.degree. C. The reaction was monitored by TLC
(DCM:MeOH=10:1). After the reaction was completed, the resulting
mixture was filtered, and the filter cake was washed with hot
methanol. The filtrate was rotary evaporated to dryness to give a
crude product (1.5 g), which was purified with silica gel column
(eluent: dichloromethane/methanol=10/1) to give the target compound
of Example 7 as a white solid (0.18 g). Yield: 93.3%. LC-MS: 358
[M+1].sup.+, t.sub.R=3.717 min. A corresponding hydrogenchloride
salt was obtained by mixing the solid with a solution of hydrogen
chloride in methanol under stirring. .sup.1H NMR (400 MHz, DMSO)
.delta. 8.30-8.16 (m, 1H), 8.09-7.96 (m, 1H), 7.87 (d, J=6.7 Hz,
2H), 7.79-7.64 (m, 1H), 7.34 (d, J=8.4 Hz, 1H), 5.38-4.14 (br, 3H),
3.74-3.16 (m, 5H), 3.05-2.65 (m, 4H), 1.91-1.60 (m, 2H).
Preparation Example 6
[0209] Synthesis of Intermediate 62
##STR00054##
[0210] Under the protection of nitrogen, 0.928 g (5.04 mmol) of
intermediate 61 was dissolved in 3.56 g (60.48 mmol, 85%) of
hydrazine hydrate, and reacted at 85.degree. C. for 2 h. The
reaction was monitored by TLC. After the reaction was completed,
the resulting mixture was cooled to room temperature, filtered, and
pumped to dryness to give a white solid (0.369 g). Yield:
39.68%.
[0211] Synthesis of Intermediate 63
##STR00055##
[0212] 0.47 g (1.34 mmol) of intermediate 9-1 was dissolved in 20
ml of dichloromethane, added with 3 drops of N,N-dimethylformamide
and cooled to 0.degree. C. 0.34 ml (4 mmol) of oxalyl chloride was
added dropwise to the mixture, then naturally warmed to room
temperature and reacted for 1 h. After the reaction was completed,
the reaction solution was concentrated and added with 20 ml of
dichloromethane, as a stock solution. 0.369 g (2 mmol) compound 62
and 0.56 ml (4 mmol) of triethylamine were dissolved in 20 ml of
dichloromethane and cooled to 0.degree. C. The solution of acyl
chloride in dichloromethane obtained above was added dropwise to
the mixture, then naturally warmed to room temperature and reacted
overnight. The reaction was monitored by TLC (PE/EA=3:1+AcOH).
After the reaction was completed, 30 ml of saturated sodium
bicarbonate solution was added and the resulting mixture was
separated into layers. The aqueous phase was extracted with
dichloromethane. The organic phases were combined, dried and rotary
evaporated to dryness to give a crude product, which was purified
with silica gel column (eluent: petroleum ether/ethyl acetate=1/1,
v/v) to give a product (0.302 g) as a yellow solid. Yield: 43.28%.
LC-MS: 518 [M+1].sup.+, t.sub.R=5.462 min.
[0213] Synthesis of Intermediate 64
##STR00056##
[0214] 0.302 g (0.58 mmol) of intermediate 63 and 0.24 ml (1.74
mmol) of triethylamine were dissolved in 10 ml of acetonitrile,
cooled to 0.degree. C., added with 0.166 g (0.87 mmol) of 4-toluene
sulfonyl chloride, and stirred overnight at room temperature. The
reaction was monitored by TLC (PE/EA=1:1). After the reaction was
completed, water was added and the resulting mixture was separated
into layers. The aqueous phase was extracted with dichloromethane.
The organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=1/1, v/v) to give a
product (0.332 g) as a yellow solid. Yield: 100%.
[0215] Synthesis of Intermediate 65
##STR00057##
[0216] 0.332 g (0.58 mmol) of intermediate 64 was dissolved in 1.25
ml of hydrochloric acid (1 M) and 10 ml of methanol, and reacted
for 30 min at 80.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated by rotary evaporation, added
with 10 ml of saturated sodium bicarbonate solution and 10 ml of
dichloromethane, and separated into layers. The aqueous phase was
extracted with dichloromethane. The organic phases were combined,
dried and rotary evaporated to dryness to give a crude product,
which was purified with silica gel column (eluent: petroleum
ether/ethyl acetate=1/1+1% Et.sub.3N, v/v) to give a product (0.214
g) as a yellow solid. Yield: 81.03%.
Example 8
(S)-(2-amino-7-(5-(3-fluoro-4-methoxyphenyl)-1,3,4-oxadiazol-2-y-
l)-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00058##
[0218] 0.214 g (0.47 mmol) of intermediate 65 was dissolved in 10
ml of methanol, added with 2 drops of concentrated hydrochloric
acid, purged with nitrogen to remove air, followed by the addition
of 0.04 g (20% m/m) of palladium 10% on carbon, purged with
hydrogen, and reacted for 4 h at 95.degree. C. The reaction was
monitored by TLC (DCM:MeOH=10:1). After the reaction was completed,
the resulting mixture was filtered, and the filter cake was washed
with plenty of methanol. The filtrate was concentrated by rotary
evaporation, added with 10 ml of saturated sodium bicarbonate
solution and 10 ml of ethyl acetate, stirred, allowed to stand and
separated into layers. The aqueous phase was extracted with ethyl
acetate. The organic phases were combined, dried, filtered and
rotary evaporated to give a crude product, which was purified with
silica gel column (eluent: DCM:MeOH=10/1, V/V) to give the target
compound of Example 8 as a faint yellow solid (0.138 g). Yield:
79.49%. LC-MS: 370 [M+1].sup.+, t.sub.R=1.334 min. A corresponding
hydrogenchloride salt was obtained by mixing the solid with a
solution of hydrogen chloride in methanol under stirring.
Preparation Example 7
[0219] Synthesis of Intermediate 69
##STR00059##
[0220] 1.0 g (5.6 mmol) of intermediate 68 was dissolved in 10 ml
of N,N-dimethylformamide, added with 2.34 ml (16.8 mmol) of
triethylamine, cooled to 0.degree. C., then added with 2.51 g (6.16
mmol) of N,N,N',N'-tetramethyl-o-(7-azabenzotriazol-1-yl)uronium
hexafluorophosphate, and stirred for 30 min. The resulting solution
was added dropwise to a solution of 2.8 g (56 mmol) of hydrazine
hydrate in 10 ml of N,N-dimethylformamide and reacted overnight.
The reaction was monitored by TLC. After the reaction was
completed, the reaction solution was rotary evaporated to dryness,
added with 10 ml of saturated sodium bicarbonate solution and 10 ml
of dichloromethane, and separated into layers. The aqueous phase
was extracted with dichloromethane. The organic phases were
combined, dried and rotary evaporated to dryness to give a crude
product, which was purified with silica gel column (eluent:
DCM/MeOH=10/1, v/v) to give a product (0.24 g) as a white solid.
Yield: 22.18%. LC-MS: 194 [M+1].sup.+, t.sub.R=1.435 min.
[0221] Synthesis of Intermediate 70
##STR00060##
[0222] 2.1 g (5.97 mmol) of intermediate 9-1 was dissolved in 80 ml
of dichloromethane, added with 10 drops of N,N-dimethylformamide
and cooled to 0.degree. C. 1.52 ml (17.92 mmol) of oxalyl chloride
was added dropwise to the mixture, then naturally warmed to room
temperature and reacted for 1 h. After the reaction was completed,
the reaction solution was concentrated and added with 80 ml of
dichloromethane, as a stock solution. 1.732 g (8.96 mmol) compound
69 and 2.5 ml (17.92 mmol) of triethylamine were dissolved in 80 ml
of dichloromethane and cooled to 0.degree. C. The solution of acyl
chloride in dichloromethane obtained above was added dropwise to
the mixture, then naturally warmed to room temperature and reacted
overnight. The reaction was monitored by TLC (PE/EA=3:1+AcOH).
After the reaction was completed, 120 ml of saturated sodium
bicarbonate solution was added and the resulting mixture was
separated into layers. The aqueous phase was extracted with
dichloromethane. The organic phases were combined, dried and
evaporated to dryness to give a crude product, which was purified
with silica gel column (eluent: petroleum ether/ethyl acetate=1/1,
v/v) to give a product (2.0 g) as a yellow solid. Yield: 63.16%.
LC-MS: 527 [M+1].sup.+, t.sub.R=2.457 min.
[0223] Synthesis of Intermediate 71
##STR00061##
[0224] 2.0 g (3.8 mmol) of intermediate 70 and 1.59 ml (11.4 mmol)
of triethylamine were dissolved in 50 ml of acetonitrile, cooled to
0.degree. C., added with 1.087 g (5.7 mmol) of 4-toluene sulfonyl
chloride, and stirred overnight at room temperature. The reaction
was monitored by TLC (PE/EA=1:1). After the reaction was completed,
water was added to the resulting mixture, which was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and evaporated to dryness to
give a crude product, which was purified with silica gel column
(eluent: petroleum ether/ethyl acetate=5/1, v/v) to give a product
(2.2 g) as a yellow solid. Yield: 100%.
[0225] Synthesis of Intermediate 72
##STR00062##
[0226] 2.2 g (3.8 mmol) of intermediate 71 was dissolved in 8.17 ml
of hydrochloric acid (1 M) and 50 ml of methanol, and reacted for
30 min at 80.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated by rotary evaporation, added
with 50 ml of saturated sodium bicarbonate solution and 50 ml of
dichloromethane, and separated into layers. The aqueous phase was
extracted with dichloromethane. The organic phases were combined,
dried and evaporated to dryness to give a crude product, which was
purified with silica gel column (eluent: petroleum ether/ethyl
acetate=1/1+1% Et.sub.3N, v/v) to give a product (0.8 g) as a
yellow solid. Yield: 45.0%.
Example 9
(S)--N-(4-(5-(7-amino-7-(hydroxymethyl)-5,6,7,8-tetrahydronaphth-
alen-2-yl)-1,3,4-oxadiazol-2-yl)phenyl)acetamide
##STR00063##
[0228] 0.8 g (1.71 mmol) of intermediate 72 was dissolved in 30 ml
of methanol, added with 5 drops of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of
0.16 g (20% m/m) of palladium 10% on carbon, purged with hydrogen,
and reacted for 4 h at 95.degree. C. The reaction was monitored by
TLC (DCM:MeOH=10:1). After the reaction was completed, the
resulting mixture was filtered, and the filter cake was washed with
plenty of methanol. The filtrate was concentrated by rotary
evaporation, added with 30 ml of saturated sodium bicarbonate
solution and 30 ml of ethyl acetate, stirred, allowed to stand and
separated into layers. The aqueous phase was extracted with ethyl
acetate. The organic phases were combined, dried, filtered and
rotary evaporated to give a crude product, which was purified with
silica gel column (eluent: DCM:MeOH=10/1, V/V) to give the target
compound of Example 9 as a faint yellow solid (0.326 g). Yield:
50.38%. LC-MS: 379 [M+1].sup.+, t.sub.R=1.103 min. A corresponding
hydrogenchloride salt was obtained by mixing the solid with a
solution of hydrogen chloride in methanol under stirring.
Preparation Example 8
[0229] Synthesis of Intermediate 76
##STR00064##
[0230] 1.0 g (4.54 mmol) of material 75, 1.9 g (13.62 mmol) of
potassium carbonate and 2.6 ml (27.00 mmol) of benzyl chloride were
suspended in 40 ml of acetone, heated at reflux and reacted
overnight. The reaction was monitored by TLC (PE/EA=3:1). After the
material 75 was reacted completely, the reaction solution was
concentrated by rotary evaporation, added with 40 ml of saturated
sodium bicarbonate solution and 40 ml of dichloromethane, stirred,
allowed to stand and separated into layers. The aqueous phase was
extracted with 40 ml of dichloromethane. The organic phases were
combined, dried and rotary evaporated to dryness to give a crude
product, which was purified with silica gel column (eluent:
petroleum ether/ethyl acetate=20/1, v/v) to give a product (1.4 g)
as a white solid. Yield: 100%.
[0231] Synthesis of Intermediate 77
##STR00065##
[0232] Under the protection of nitrogen, 1.4 g (4.54 mmol) of
intermediate 76 was suspended in 40 ml of methanol and 6.4 ml
(108.96 mmol, 85%) of hydrazine hydrate, and reacted at 85.degree.
C. for 180 min. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated by rotary evaporation, cooled to
room temperature, added with 20 ml of water and stirred to
precipitate out a solid, filtered, washed with water, and pumped to
dryness to give a product (1.4 g), as a white solid. Yield: 100%.
LC-MS: 311 [M+1].sup.+, t.sub.R=1.799 min.
[0233] Synthesis of Intermediate 78
##STR00066##
[0234] 1.36 g (3.87 mmol) of intermediate 9-1 was dissolved in 27
ml of dichloromethane, added with 0.01 g (cat.) of
N,N-dimethylformamide and cooled to 0.degree. C. 1.0 ml (11.61
mmol) of oxalyl chloride was added dropwise to the mixture, then
naturally warmed to room temperature and reacted for 1 h. After the
reaction was completed, the reaction solution was concentrated and
added with 30 ml of dichloromethane, as a stock solution. 1.2 g
(3.87 mmol) of intermediate 77 and 1.62 ml (11.61 mmol) of
triethylamine were dissolved in 30 ml of dichloromethane and cooled
to 0.degree. C. The solution of acyl chloride in dichloromethane
obtained above was added dropwise to the mixture, then naturally
warmed to room temperature and reacted overnight. The reaction was
monitored by TLC (PE/EA=3:1+AcOH). After the reaction was
completed, 60 ml of saturated sodium bicarbonate solution was added
and the resulting mixture was separated into layers. The aqueous
phase was extracted with dichloromethane. The organic phases were
combined, dried and rotary evaporated to dryness to give a crude
product, which was purified with silica gel column (eluent:
petroleum ether/ethyl acetate=2/1, v/v) to give a product (1.4 g)
as a yellow solid. Yield: 56.2%. LC-MS: 644 [M+1].sup.+,
t.sub.R=2.396 min.
[0235] Synthesis of Intermediate 79
##STR00067##
[0236] 1.4 g (2.17 mmol) of intermediate 78 and 1.0 ml (6.51 mmol)
of triethylamine were dissolved in 56 ml of acetonitrile, cooled to
0.degree. C., added with 0.5 g (2.61 mmol) of 4-toluene sulfonyl
chloride, and stirred overnight at room temperature. The reaction
was monitored by TLC (PE/EA=1:1). After the reaction was completed,
water was added to the resulting mixture, which was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=3/1, v/v) to give a
product (0.6 g) as a faint yellow solid. Yield: 44.2%.
[0237] Synthesis of Intermediate 80
##STR00068##
[0238] 0.6 g (0.95 mmol) of intermediate 79 was dissolved in 2.4 ml
of hydrochloric acid (1 M) and 30 ml of methanol, and reacted for
120 min at 80.degree. C. to precipitate out a solid. The reaction
was monitored by TLC (PE/EA=1:1+Et.sub.3N). After the reaction was
completed, the resulting mixture was filtrated under vacuum. The
mother liquor was concentrated by rotary evaporation, added with 30
ml of saturated sodium bicarbonate solution and 30 ml of
dichloromethane, and separated into layers. The aqueous phase was
extracted with dichloromethane. The organic phases were combined,
dried and rotary evaporated to dryness to give a crude product,
which was purified with silica gel column (eluent: petroleum
ether/ethyl acetate=1/2+1% Et.sub.3N, v/v) to give a product (0.426
g) as a white solid. Yield: 76.57%.
Example 10
(S)-4-(5-(7-amino-7-(hydroxymethyl)-5,6,7,8-tetrahydronaphthale-
n-2-yl)-1,3,4-oxadiazol-2-yl)-2-(trifluoromethyl)phenol
##STR00069##
[0240] 0.426 g (0.727 mmol) of intermediate 80 was dissolved in 16
ml of methanol, added with 0.01 ml of concentrated hydrochloric
acid, purged with nitrogen to remove air, followed by the addition
of 0.09 g of palladium 10% on carbon, purged with hydrogen, and
reacted overnight at 95.degree. C. The reaction was monitored by
TLC (DCM:MeOH=10:1). After the reaction was completed, the
resulting mixture was filtered, and the filter cake was washed with
hot methanol. The filtrate was concentrated by rotary evaporation,
added with 1% sodium hydrate solution to adjust pH to 9-10. The
aqueous phase was extracted with dichloromethane, added with
hydrochloric acid (1 M) to adjust pH 5.8-6.2 to precipitate out a
white solid, stirred for 30 min at room temperature, and then
filtered under vacuum to give the target compound of Example 10 (25
mg) as a white solid, which is prone to oxidative degradation and
stored at low temperature under the protection of nitrogen. Yield:
8.5%. LC-MS: 406 [M+1].sup.+, t.sub.R=1.328 min. A corresponding
hydrogenchloride salt was obtained by mixing the solid with a
solution of hydrogen chloride in methanol under stirring.
Preparation Example 9
[0241] Synthesis of Intermediate 84
##STR00070##
[0242] 1.0 ml (14.4 mmol) of thionyl chloride was added to 20 ml of
methanol at low temperature, stirred and reacted for 1 h in an
ice-water bath, followed by the addition of 1.0 g of material 83 to
the reaction vessel, naturally warmed, then heated to 55.degree.
C., stirred and reacted for 1 h. The reaction was monitored by TLC
(PE/EA=3:1+AcOH). After the material 83 was reacted completely, the
reaction solution was concentrated by rotary evaporation, added
with 20 ml of saturated sodium bicarbonate solution and 20 ml of
ethyl acetate, stirred, allowed to stand and separated into layers.
The aqueous phase was extracted with 40 ml of ethyl acetate twice.
The organic phases were combined, dried and rotary evaporated to
dryness to give a crude product (0.7 g), as an oily substance.
Yield: 65.7%.
[0243] Synthesis of Intermediate 85
##STR00071##
[0244] Under the protection of nitrogen, 0.7 g (3.2 mmol) of
intermediate 84 was suspended in 7 ml of methanol and 1.9 ml (32
mmol, 85%) of hydrazine hydrate, and reacted at room temperature
overnight. The reaction was monitored by TLC (PE/EA=3:1). After the
reaction was completed, the resulting mixture was added with 20 ml
of water and 20 ml of DCM, stirred, separated and extracted with
DCM twice. The organic phases were combined, dried, filtered,
concentrated under reduced pressure and pumped to dryness to give a
white solid (0.7 g). Yield: 100%. LC-MS: 223 [M+1].sup.+,
t.sub.R=1.879 min.
[0245] Synthesis of Intermediate 86
##STR00072##
[0246] 1.0 g (3.1 mmol) of intermediate 9-1 was dissolved in 40 ml
of dichloromethane, added with 0.01 g (cat.) of
N,N-dimethylformamide and cooled to 0.degree. C. 0.8 ml (9.3 mmol)
of oxalyl chloride was added dropwise to the mixture, then
naturally warmed to room temperature and reacted for 1 h. After the
reaction was completed, the reaction solution was concentrated and
added with 20 ml of dichloromethane, as a stock solution. 0.7 g
(3.87 mmol) intermediate 85 and 1.3 ml (9.3 mmol) of triethylamine
were dissolved in 20 ml of dichloromethane and cooled to 0.degree.
C. The solution of acyl chloride in dichloromethane obtained above
was added dropwise to the mixture, then naturally warmed to room
temperature and reacted overnight. The reaction was monitored by
TLC (PE/EA=3:1+AcOH). After the reaction was completed, 40 ml of
saturated sodium bicarbonate solution was added and the resulting
mixture was separated into layers. The aqueous phase was extracted
with dichloromethane. The organic phases were combined, dried and
rotary evaporated to dryness to give a crude product (1.5 g).
Yield: 87.1%.
[0247] Synthesis of Intermediate 87
##STR00073##
[0248] 1.5 g (3.1 mmol) of intermediate 86 and 1.3 ml (9.3 mmol) of
triethylamine were dissolved in 60 ml of acetonitrile, cooled to
0.degree. C., added with 0.88 g (4.65 mmol) of 4-toluene sulfonyl
chloride, and stirred overnight at room temperature. The reaction
was monitored by TLC (PE/EA=1:1). After the reaction was completed,
water was added and the resulting mixture was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=3/1, v/v) to give a
product (1.3 g) as a faint yellow solid. Yield: 78.0%. LC-MS: 538
[M+1].sup.+, t.sub.R=3.251 min.
[0249] Synthesis of Intermediate 88
##STR00074##
[0250] 1.3 g (2.4 mmol) of intermediate 87 was dissolved in 6 ml of
hydrochloric acid (1 M) and 40 ml of methanol, and reacted for 3 h
at 80.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated under reduced pressure, added
with sodium hydrate solution (10%) and saturated sodium bicarbonate
solution to adjust pH about 8, and extracted with dichloromethane.
The organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=1/1+1% Et.sub.3N,
v/v) to give a product (0.6 g) as a white solid. Yield: 50.2%.
Example 11
(S)-(2-amino-7-(5-(3-fluoro-4-(trifluoromethyl)phenyl)-1,3,4-ox-
adiazol-2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00075##
[0252] 0.6 g (1.2 mmol) of intermediate 88 was dissolved in 20 ml
of methanol, added with 0.01 ml of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of
0.12 g (20% m/m) of 10% wet palladium on carbon, purged with
hydrogen, and reacted for 6 h at 85.degree. C. The reaction was
monitored by TLC (DCM:MeOH=10:1). After the reaction was completed,
the resulting mixture was filtered while hot, and the filter cake
was washed with hot methanol. The filtrate was concentrated by
rotary evaporation, added with 6 ml of methanol and a solution (1
ml, 10%) of hydrogen chloride in methanol, warmed to reflux,
stirred for 30 min, then naturally cooled to room temperature, and
filtered under vacuum to give a white solid (0.1 g). The filtrate
was rotary evaporated to dryness and the processes above-mentioned
were repeated to give a white solid (0.076 g). The two white solids
obtained were combined to give the target compound of Example 11
(0.176 g, hydrogenchloride salt). Yield: 33.0%. LC-MS: 408
[M+1].sup.+, t.sub.R=1.936 min.
Preparation Example 10
[0253] Synthesis of Intermediate 92
##STR00076##
[0254] 1.7 ml (23.1 mmol) of thionyl chloride was added to 40 ml of
methanol at low temperature, stirred and reacted for 1 h in an
ice-water bath, followed by the addition of 2.0 g of material 91,
naturally warmed, then heated to 80.degree. C., stirred and reacted
for 1 h. The reaction was monitored by TLC (PE/EA=3:1+AcOH). After
the material 91 was reacted completely, the reaction solution was
concentrated by rotary evaporation, added with 20 ml of saturated
sodium bicarbonate solution and 20 ml of ethyl acetate, stirred,
allowed to stand and separated into layers. The aqueous phase was
extracted with 40 ml of ethyl acetate twice. The organic phases
were combined, dried and rotary evaporated to dryness to give a
crude product (1.5 g), as an oily substance. Yield: 71.6%.
[0255] Synthesis of Intermediate 93
##STR00077##
[0256] Under the protection of nitrogen, 1.5 g (5.5 mmol) of
intermediate 92 was suspended in 30 ml of methanol and 3.2 ml (55
mmol, 85%) of hydrazine hydrate, and reacted at room temperature
overnight. The reaction was monitored by TLC (PE/EA=3:1). After the
reaction was completed, the resulting mixture was added with 20 ml
of water and 20 ml of DCM, stirred, separated and extracted with
DCM twice. The organic phases were combined, dried, filtered,
concentrated under reduced pressure and pumped to dryness to give a
white solid (1.5 g). Yield: 100%. LC-MS: 273 [M+1].sup.+,
t.sub.R=2.034 min.
[0257] Synthesis of Intermediate 94
##STR00078##
[0258] 1.7 g (5 mmol) of intermediate 9-1 was dissolved in 68 ml of
dichloromethane, added with 0.02 g (cat.) of N,N-dimethylformamide
and cooled to 0.degree. C. 1.3 ml (15 mmol) of oxalyl chloride was
added dropwise to the mixture, then naturally warmed to room
temperature and reacted for 1 hour. After the reaction was
completed, the reaction solution was concentrated and added with 34
ml of dichloromethane, as a stock solution. 1.5 g (5.5 mmol)
intermediate 93 and 2.1 ml (15 mmol) of triethylamine were
dissolved in 40 ml of dichloromethane and cooled to 0.degree. C.
The solution of acyl chloride in dichloromethane obtained above was
added dropwise to the mixture, then naturally warmed to room
temperature and reacted overnight. The reaction was monitored by
TLC (PE/EA=3:1+AcOH). After the reaction was completed, 40 ml of
saturated sodium bicarbonate solution was added and the resulting
mixture was separated into layers. The aqueous phase was extracted
with dichloromethane. The organic phases were combined, dried and
rotary evaporated to dryness to give a crude product (3.0 g).
Yield: 100%.
[0259] Synthesis of Intermediate 95
##STR00079##
[0260] 3.0 g (5 mmol) of crude intermediate 94 and 2.1 ml (15 mmol)
of triethylamine were dissolved in 120 ml of acetonitrile, cooled
to 0.degree. C., added with 1.1 g (6 mmol) of 4-toluene sulfonyl
chloride, and stirred overnight at room temperature. The reaction
was monitored by TLC (PE/EA=1:1). After the reaction was completed,
water was added and the resulting mixture was separated into
layers. The aqueous phase was extracted with dichloromethane. The
organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=10/1, v/v) to give a
product (1.3 g) as a bubble-shaped solid. Yield: 44.2%. LC-MS:
588.5 [M+1].sup.+, t.sub.R=3.300 min.
[0261] Synthesis of Intermediate 96
##STR00080##
[0262] 1.3 g (2.2 mmol) of intermediate 95 was dissolved in 5.5 ml
of hydrochloric acid (1 M) and 50 ml of methanol, and reacted for 2
h at 90.degree. C. The reaction was monitored by TLC
(PE/EA=1:1+Et.sub.3N). After the reaction was completed, the
resulting mixture was concentrated under reduced pressure, added
with sodium hydrate solution (10%) and saturated sodium bicarbonate
solution to adjust pH about 8, and extracted with dichloromethane.
The organic phases were combined, dried and rotary evaporated to
dryness to give a crude product, which was purified with silica gel
column (eluent: petroleum ether/ethyl acetate=1/1+1% Et.sub.3N,
v/v) to give a product (0.6 g) as a white solid. Yield: 49.8%.
Example 12
(S)-(2-amino-7-(5-(3,5-bis(trifluoromethyl)phenyl)-1,3,4-oxadia-
zol-2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00081##
[0264] 0.6 g (1.1 mmol) of intermediate 96 was dissolved in 34 ml
of methanol, added with 0.01 ml of concentrated hydrochloric acid,
purged with nitrogen to remove air, followed by the addition of
0.12 g (20% m/m) of 10% wet palladium on carbon, purged with
hydrogen, and reacted for 6 h at 90.degree. C. The reaction was
monitored by TLC (DCM:MeOH=10:1). After the reaction was completed,
the resulting mixture was filtered while hot, and the filter cake
was washed with hot methanol. The filtrate was concentrated by
rotary evaporation to give a crude product as a white solid. The
crude product was purified with silica gel column (eluent:
dichloromethane/methanol=5/1, v/v), then dissolved in 20 ml of DCM
(containing a small amount of methanol) and 20 ml of water, stirred
and separated. The aqueous phase was extracted with DCM. The DCM
phases were combined, dried, and filtered. The filtrate was added
with an appropriate amount of solution of hydrogen chloride in
methanol, rotary evaporated to dryness, and pumped to dryness under
reduced pressure to give the target compound of Example 12 (0.126
g, hydrogenchloride salt). Yield: 33%. Purity: 96.1%. LC-MS: 458
[M+1].sup.+, t.sub.R=1.961 min.
Example 13
(S)-(2-amino-7-(5-(3,4-diethoxyphenyl)-1,3,4-oxadiazol-2-yl)-1,-
2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00082##
[0266] The chiral compound of the present example was obtained by
using the chiral intermediate 7-1 as a starting material in a
manner similar to the preparation method of example 20 in
WO2013181840. LC-MS: 410.2 [M+1].sup.+, t.sub.R=1.757 min. H NMR
(400 MHz, DMSO) .delta. 7.90-7.77 (m, 2H), 7.68 (dd, J=8.4, 1.9 Hz,
1H), 7.59 (d, J=1.9 Hz, 1H), 7.33 (d, J=7.9 Hz, 1H), 7.17 (d, J=8.5
Hz, 1H), 4.98 (br s, 1H), 4.14 (p, J=6.8 Hz, 5H), 3.29 (s, 2H),
3.04-2.60 (m, 5H), 1.88-1.59 (m, 2H), 1.43-1.30 (m, 6H). A
hydrogenchloride salt was obtained by processing the chiral
compound with a small amount of solution of hydrogen chloride in
methanol.
Example 14
(S)-(2-amino-7-(5-(4-isopropoxy-3-(trifluoromethyl)phenyl)-1,3,-
4-oxadiazol-2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methanol
##STR00083##
[0268] The chiral compound of the present example was obtained by
using the chiral intermediate 7-1 as a starting material in a
manner similar to the preparation method of example 11 in
WO2013181840. A hydrogenchloride salt was obtained by processing
the chiral compound with a small amount of solution of hydrogen
chloride in methanol. LC-MS: 448.5 [M+1].sup.+, t.sub.R=3.183 min.
H NMR (400 MHz, DMSO) .delta. 8.43-8.31 (m, 1H), 8.27 (s, 1H), 8.13
(s, 3H), 7.94 (d, J=5.5 Hz, 2H), 7.56 (d, J=9.0 Hz, 1H), 7.40 (d,
J=8.6 Hz, 1H), 5.61 (t, J=5.1 Hz, 1H), 5.03-4.87 (m, 1H), 3.47 (d,
J=5.0 Hz, 2H), 3.17-2.79 (m, 4H), 2.07-1.86 (m, 2H), 1.35 (d, J=6.0
Hz, 6H).
[0269] Biological Activity Assays
[0270] The compounds of the invention were detected for biological
activity hereinafter:
Example 15 Effects of Amino Alcohol Compounds on the Expression of
CD4, CD8 and CD19 on Peripheral Blood Cell in Mice
[0271] 1. Test Materials:
[0272] Mice (C57 BL/6, 8 weeks)
[0273] FITC Rat Anti-Mouse CD8a: BD, Cat. #553030.
[0274] PE Rat Anti-Mouse CD4: BD, Cat. #557308.
[0275] APC Rat Anti-Mouse CD19: BD, Cat. #561738.
[0276] The compounds used in the test were prepared by the chemical
department of Beijing Forelandpharma Co. Ltd.
[0277] 2. Test Method:
[0278] Routs of administration: intragastric administration, once a
day, and continuous administration for four days.
[0279] After the administration, supraorbital venous blood was
collected from the mouse, added to an EP tube (1.5 ml) containing
an anticoagulant, and kept on the ice. The resulting mixture was
transferred to a test tube, centrifuged at 4.degree. C. and 1200
rpm/min for 5 min, removed with the supernatant liquid, added with
a lysate, processed with cell lysis on the ice for 5 min, then
pre-dyed, added with diluted antibody, and incubated away from
light for 30 min. After the incubation, the resulting mixture was
washed, removed with the supernatant liquid, added with stationary
liquid, and kept in a refrigerator at 4.degree. C. away from light
for the test of flow cytometry.
[0280] 3. Test Results
TABLE-US-00003 TABLE 1 Effects of Amino Alcohol Compounds on the
Expression of CD4, CD8 and CD19 on Peripheral Blood Cell in Mice
Num- Exam- CD4 (%) CD8 (%) CD19 (%) ber ple 1 mg/kg 5 mg/kg 1 mg/kg
5 mg/kg 1 mg/kg 5 mg/kg 1 vehicle 25.7 14.5 47.5 2 13 23.4 11.6
12.7 9.4 56.8 53.8 3 1 10.9 1.2 10.0 1.8 39.0 17.5 4 2 23.1 16.4
13.0 8.6 44.8 35.2 5 3 22.1 7.2 11.9 6.1 39.3 24.7 6 6 17.8 8.7
17.0 5.7 34.0 29.1 7 4 27.1 27.0 14.5 12.3 47.1 43.6 8 7 26.5 13.4
14.5 9.2 45.2 40.6 9 8 33.4 30.0 16.6 16.0 36.8 41.7 10 9 33.1 31.9
17.3 17.0 42.5 44.3 11 5 31.5 27.1 16.8 14.7 44.3 43.8 12 10 31.1
27.7 15.3 14.0 44.3 35.7 13 11 23.5 20.5 14.9 13.8 52.1 54.0 14 12
15.2 2.8 9.6 3.4 53.2 38.1
Example 16 .beta.-arrestin Testing Experiments (PathHunter
.beta.-Arrestin Testing System)
[0281] Test Method
[0282] 1. PathHunter .beta.-arrestin testing system was used to
detect the biological activity of the compounds.
[0283] 2. .beta.-arrestin engineering cells were cultured on
386-well culture plates and placed in an incubator at 37.degree.
C.
[0284] 3. The sample to be tested was diluted with reaction
solution in 5-fold dilution.
[0285] 4. The diluted sample to be tested was added to the
engineering cells and the reaction was induced and observed.
[0286] 5. The chemiluminescence signal produced in the test may be
detected by multifunctional enzyme marker (PerkinElmer
Envision.TM.).
[0287] Test Analysis and Results
[0288] 6. The data obtained was analyzed using the data analysis
software of CBIS (ChemInnovation, CA) to achieve EC.sub.50, and the
test results are shown in Table 2.
TABLE-US-00004 TABLE 2 Biological Activity of the Compounds in
Example 3 and Example 13 .beta.-arrestin Test EC.sub.50 (nM)
Compound S1P1 S1P2 S1P3 S1P4 S1P 25.2 26.4 26.3 189.7 Example 13
21.6 >100,000 613 >100,000 Example 3 4.56 >100,000 2114
>100,000
Example 17 Test of Fluorescence Detection (FLIPR Assay)
[0289] Test Method and Results
[0290] The sample to be tested was dissolved in DMSO and diluted
with detection buffer solution in 3-fold dilution. A reagent and a
positive control were diluted in the same way.
[0291] The reactions of the agonist, the reagent and the positive
control were detected by a device of FLIPRTETRA with a total
detection time of 180 s to estimate the ability to activate GPCR
(S1P5) of each compound. The results are shown in Table 3.
TABLE-US-00005 TABLE 3 Biological Activity of the Compounds in
Example 3 and Example 13 FLIPR Test EC.sub.50 (nM) Compound S1P5
S1P 23 Example 13 >10,000 Example 3 250
Example 18
[0292] Test Method
[0293] 1. HEK293 engineering cells stably expressed with hERG
potassium channel were used to test the compounds.
[0294] 2. Patch Clamp Detection
[0295] The cells were separated by TrypLE.TM. Express before the
test. 3.times.10.sup.3 of cells were spread on a cover plate,
cultured in 24-well plate and tested 18 hours later. The signals
produced by voltage stimulation of potassium currents in cells were
recorded by electrophysiological techniques.
[0296] Data Analysis and Results
[0297] First of all, currents after the action of drug of each
concentration and blank control were standardized separately
( Peak .times. .times. tail .times. .times. current .times. .times.
compound Peak .times. .times. tail .times. .times. current .times.
.times. vehicle ) , ##EQU00001##
and then the corresponding inhibition ratio of each concentration
was calculated
( 1 .times. - .times. Peak .times. .times. tail .times. .times.
current .times. .times. compound Peak .times. .times. tail .times.
.times. current .times. .times. vehicle ) . ##EQU00002##
The average and the standard error were calculated for each
concentration, and 50% inhibiting concentration of each compound
was calculated according to the following equation:
inhibition = 1 1 + ( IC .times. .times. 50 C ) h ##EQU00003##
[0298] The dose-dependent effect was obtained through nonlinear
fitting with the equation above. C refers to drug concentration,
IC50 refers to 50% inhibiting concentration and h refers to hill
coefficient. The curve fitting and the calculation of IC50 were
processed by IGOR software. The results are shown in Table 4.
TABLE-US-00006 TABLE 4 HERG Test of Example 13 and Example 14 hERG
Test Compound IC.sub.50 (.mu.M) Example 13 1.2 Example 14 0.87
Example 3 7.8
[0299] The embodiments of the present invention have been described
above. However, the present invention is not limited to the
embodiments above mentioned. Any modification, equivalent
substitution and improvement, etc., which are made within the
spirit and principle of the invention, should fall within the scope
of the invention.
* * * * *